Characterization of reactive intermediates by diffusion-ordered NMR spectroscopy: A snapshot of the reaction of (CO2)-C-13 with [Cp2Zr(Cl)H] - Schlorer, NE; E. J. Cabrita; Berger, S (Angewandte Chemie-International Edition, 2002)
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Energy transfer between polyamine chains bearing naphthalene terminal units and K-3 Co(CN)(6) : an example of a molecular photoreactor - Arranz, P; Albelda, MT; Garcia-Espana, E; J. C. Lima; Lodeiro, C.; de Melo, JS; A. J. Parola; Pina, F (Journal of the Chemical Society-Dalton Transactions, 2002)
Molecular photoreactors consisting of polyamine chains (receptors) bearing terminal naphthalene units (antennae) are described. The receptors are used to bind the substrate hexacyanocobaltate(III) and the antennae to transfer energy to the complex and thus promote a photoaquation reaction.
ISI Document Delivery No.: 576AVTimes Cited: 2 Cited Reference Count: 29 Arranz, P Albelda, MT Garcia-Espana, E Lima, JC Lodeiro, C de Melo, JS Parola, AJ Pina, F Seixas de Melo, Sergio/A-5388-2008; Pina, Fernando/C-8161-2011; Albelda, M. Teresa/J-5929-2012; Lodeiro, Carlos/B-4793-2013; Parola, A. Jorge/F-4048-2010; REQUIMTE, AL/H-9106-2013; Lima, Joao/F-3658-2010 Seixas de Melo, Sergio/0000-0001-9708-5079; Lodeiro, Carlos/0000-0001-5582-5446; Parola, A. Jorge/0000-0002-1333-9076; Lima, Joao/0000-0003-0528-1967 Royal soc chemistry Cambridge

Switching from intramolecular energy transfer to intramolecular electron transfer by the action of pH and Zn2+ co-ordination - Albelda, MT; Diaz, P; Garcia-Espana, E; J. C. Lima; Lodeiro, C.; de Melo, JS; A. J. Parola; Pina, F; Soriano, C (Chemical Physics Letters, 2002)
Intramolecular electron (eT) and energy transfer (ET) have shown to occur in a covalently linked donor-acceptor (CLDA) system consisting of a naphthalene donor covalently linked through a polyamine chain connector to an anthracene acceptor; the connector has been chosen in order to switch ON or OFF the energy flux as a function of its protonation state as well as by co-ordination to Zn2+. The largest energy transfer efficiency (eta = 0.61) occurs for the fully protonated form (pH < 2), while at pH > 9 (eT) from the lone pairs of the nitrogens to the excited fluorophore takes place, leading to complete quenching of the emission. On the other hand at neutral and basic pH values. coordination of Zn2+ prevents the eT quenching allowing the ET process to occur. (C) 2002 Elsevier Science B.V. All rights reserved.
ISI Document Delivery No.: 522VVTimes Cited: 28 Cited Reference Count: 15 Albelda, MT Diaz, P Garcia-Espana, E Lima, JC Lodeiro, C de Melo, JS Parola, AJ Pina, F Soriano, C Pina, Fernando/C-8161-2011; Seixas de Melo, Sergio/A-5388-2008; Albelda, M. Teresa/J-5929-2012; Lodeiro, Carlos/B-4793-2013; Parola, A. Jorge/F-4048-2010; Lima, Joao/F-3658-2010 Seixas de Melo, Sergio/0000-0001-9708-5079; Lodeiro, Carlos/0000-0001-5582-5446; Parola, A. Jorge/0000-0002-1333-9076; Lima, Joao/0000-0003-0528-1967 Elsevier science bv Amsterdam

Acidianus ambivalens type-II NADH dehydrogenase: genetic characterisation and identification of the flavin moiety as FMN - Bandeiras, Tiago M; Salgueiro, , Carlos A.; Kletzin, Arnulf; Gomes, C.M.; Teixeira, Miguel (FEBS Letters, 2002)
The thermoacidophilic archaeon Acidianus ambivalens contains a monomeric 47 kDa type-II NADH dehydrogenase (NDH), which contains a covalently bound flavin. In this work, by a combination of several methods, namely 31P-nuclear magnetic resonance and fluorescence spectroscopies, it is proven that this enzyme contains covalent FMN, a novelty among this family of enzymes, which were so far thought to mainly have the flavin dinucleotide form. Discrimination between several possible covalent flavin linkages was achieved by spectral and fluorescence experiments, which identified an 8α-N(1)-histidylflavin-type of linkage. Analysis of the gene-deduced amino acid sequence of type-II NDH showed no transmembranar helices and allowed the definition of putative dinucleotide and quinone binding motifs. Further, it is suggested that membrane anchoring can be achieved via amphipatic helices.

Adaptive observer based fault diagnosis approach applied to a thermal plant - Palma, L. B.; Coito, F.; Silva, R. (submitted to 10th Mediterranean Conference on Control and Automation, 2002)
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Determination of the orientation of the axial ligands and of the magnetic properties of the haems in the tetrahaem ferricytochrome from Shewanella frigidimarina - Ricardo O. Louro; Miguel Pessanha; Graeme A. Reid; Stephen K. Chapman; Turner, David L.; Salgueiro, C.A. (FEBS Letters, 2002)
The unambiguous assignment of the nuclear magnetic resonance (NMR) signals of the α-substituents of the haems in the tetrahaem cytochrome isolated from Shewanella frigidimarina NCIMB400, was made using a combination of homonuclear and heteronuclear experiments. The paramagnetic 13C shifts of the nuclei directly bound to the porphyrin of each haem group were analysed in the framework of a model for the haem electronic structure. The analysis yields g-tensors for each haem, which allowed the assignment of some electron paramagnetic resonance (EPR) signals to specific haems, and the orientation of the magnetic axes relative to each haem to be established. The orientation of the axial ligands of the haems was determined semi-empirically from the NMR data, and the structural results were compared with those of the homologous tetrahaem cytochrome from Shewanella oneidensis MR-1 showing significant similarities between the two proteins.

Dielectric studies of the nematic mixture E7 on a hydroxypropylcellulose substrate - M. T. Viciosa; A. M. Nunes; A. Fernandes; P. L. Almeida; M. H. Godinho; M. D. Dionísio (Liquid Crystals, 2002)

Interictal spike EEG source analysis in hypothalamic hamartoma epilepsy - Alberto Leal; Vitorina Passão; Eulália Calado; José P Vieira; João P Cunha (Clinical Neurophysiology, 2002)

The murMN operon: a functional connection between antibiotic resistance and antibiotic tolerance in Streptococcus pneumoniae - S. R. Filipe; E. Severina; A. Tomasz (Proc. Natl. Acad. Sci. USA, 2002)

A new approach to the initial value problem - Ortigueira, M. (5th Portuguese Conference on Automatic Control. :377–382, 2002)
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O projecto OCTOPUS: O módulo Reconhecedor de Zonas Oxidadas - Oliveira, R.; Pita, H.; Coito, F.; Steiger, Garção (ISEL Lisboa, Portugal, 2002)
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Patologias no edifício lote 19 da Quinta do Barroso Norte no Barreiro - Amado, Miguel (2002)

Adaptive observer based fault diagnosis approach applied to a thermal plant - Palma, L. B.; Coito, F.; Silva, R. (submitted to 10th Mediterranean Conference on Control and Automation, 2002)
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Catalytic asymmetric dihydroxylation of olefins using a recoverable and reusable OsO42- in ionic liquid bmim PF6 - Branco, L. C.; C. A. M. Afonso (Chemical Communications, 2002)
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Times Cited: 53 Branco, Luis/C-6426-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, PHOTO/M-4229-2013; Mateus Afonso, Carlos Alberto/M-7833-2013; REQUIMTE, LAQV/N-9835-2013; iMed.ULisboa, iMed.ULisboa/C-6292-2014; iMed.ULisboa, BioOrgChem /B-4209-2014 Branco, Luis/0000-0003-2520-1151; Mateus Afonso, Carlos Alberto/0000-0002-7284-5948; 54

Color stabilization of anthocyanins: Effect of SDS micelles on the acid-base and hydration kinetics of malvidin 3-glucoside (Oenin) - J. C. Lima; Vautier-Giongo, C; Lopes, A; Melo, E; Quina, FH; Macanita, AL (Journal of Physical Chemistry a, 2002)
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Construção de um Bio-Eléctrodo Específico para Determinação de Nitritos - Almeida, M. G.; Tavares, P; Moura, J. J. G. (Bol. Soc. Port. Química, 2002)
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Crystal structure of a prostate kallikrein isolated from stallion seminal plasma: A homologue of human PSA - Carvalho, A. L.; Sanz, L.; Barettino, D.; Romero, A.; Calvete, J. J.; Romao, M. J. (Journal of Molecular Biology, 2002)
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Times Cited: 51 Carvalho, Ana Luisa/G-5638-2011; Romao, Maria/A-4115-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Carvalho, Ana Luisa/0000-0002-3824-0240; Romao, Maria/0000-0002-3004-0543; 52

Crystallization and preliminary X-ray diffraction analysis of two pH-dependent forms of a di-haem cytochrome c peroxidase from Pseudomonas nautica - Dias, João M.; Bonifácio, Cecília; Alves, Teresa; Moura, José J. G.; Moura, Isabel; Romão, Maria João (Acta Crystallographica Section D, 2002)
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Keywords: calcium-binding site; Cytochromes; di-haem cytochrome c peroxidase; Pseudomonas nautica.

Crystallization and preliminary X-ray diffraction analysis of two pH-dependent forms of a di-haem cytochrome c peroxidase from Pseudomonas nautica - Dias, J. M.; Bonifacio, C.; Alves, T.; Moura, J. J. G.; Moura, I.; Romao, M. J. (Acta Crystallographica Section D-Biological Crystallography, 2002)
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Times Cited: 4 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 4

Dynamics of linear poly(methylphenylsiloxane) by time-resolved fluorescence: Slow vs fast relaxations and low-temperature behavior in chains of different lengths - Dias, FB; J. C. Lima; Horta, A; Pierola, IF; Macanita, AL (Macromolecules, 2002)
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Fractional Discrete-Time Signal Processing: Scale Conversion and Linear Prediction - Manuel Ortigueira; Matos, C.; Piedade, M.S. (Nonlinear Dynamics, 2002)
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Gene sequence and the 1.8 angstrom crystal structure of the tungsten-containing formate dehydrogenase from Desulfolvibrio gigas - Raaijmakers, H.; Macieira, S.; Dias, J. M.; Teixeira, S.; Bursakov, S.; Huber, R.; Moura, J. J. G.; Moura, I.; Romao, M. J. (Structure, 2002)
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Times Cited: 66 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, AL/H-9106-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 72

Ground and excited state properties of polyamine chains bearing two terminal naphthalene units - de Melo, JS; Albelda, MT; Diaz, P; Garcia-Espana, E; Lodeiro, C.; Alves, S; J. C. Lima; Pina, F; Soriano, C (Journal of the Chemical Society-Perkin Transactions 2, 2002)
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Highly selective transport of organic compounds by using supported liquid membranes based on ionic liquids - Branco, L. C.; Crespo, J. G.; C. A. M. Afonso (Angewandte Chemie-International Edition, 2002)
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Times Cited: 143 Branco, Luis/C-6426-2013; Crespo, Joao/D-8719-2013; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, CENG/M-4555-2013; Mateus Afonso, Carlos Alberto/M-7833-2013; REQUIMTE, LAQV/N-9835-2013; iMed.ULisboa, iMed.ULisboa/C-6292-2014; iMed.ULisboa, BioOrgChem /B-4209-2014 Branco, Luis/0000-0003-2520-1151; Mateus Afonso, Carlos Alberto/0000-0002-7284-5948; 148

Impactos sociais do desinvestimento [Social Impacts of divestment] - António Moniz; Gomes, Cláudia (University Library of Munich, Germany, 2002)
The resulting economic integration of industrial processes and manufacturing internationalisation lead several authors to argue that world economy is globalised. In this context, the approach to the divestment concept without an social and económical context, does not show a group of associated practices and representations. Choices and options are motivated by exogenous forces that pushes companies to determine strategies that stop capital investment on new equipment goods, or on other imaterial goods. This type of strategy is designated by "divestment". The social level of consequencies are not due to the closing down or de-localization of production units that are divesting, but can be materialised of efects that are irreversible. This means unemployment, de-skilling, labour precarization and even emergence of new forms of social exclusion in former industrialised regions.
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Keywords: di-vestment; globalization; labour markets; strategical management

Intramolecular addition of acyldiazenecarboxylates onto double bonds in the synthesis of heterocycles - Prata, JV; Clemente, DTS; Prabhakar, S.; A. M. Lobo; Mourato, I; Branco, P. S. (JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 1, 2002)
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Long range electron transfer quenching in polyamine chains bearing a terminal naphthalene unit - Pina, F; J. C. Lima; Lodeiro, C.; de Melo, JS; Diaz, P; Albelda, MT; Garcia-Espana, E (Journal of Physical Chemistry a, 2002)
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Manipulation of the reactivity of a synthetic anthocyanin analogue in aqueous micellar media - Vautier-Giongo, C; Yihwa, C; Moreira, PF; J. C. Lima; Freitas, AA; Alves, M; Quina, FH; Macanita, AL (Langmuir, 2002)
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Micelle effects on multi state/multifunctional systems based on photochromic flavylium compounds. The case of luteolinidin - Melo, MJ; Moura, S.; Maestri, M; Pina, F (Journal of Molecular Structure, 2002)
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Times Cited: 10 Melo, Maria Joao/C-8594-2011; Pina, Fernando/C-8161-2011; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Melo, Maria Joao/0000-0001-7393-6801; Pina, Fernando/0000-0001-8529-6848; Si 10

Molybdenum enzymes in reactions involving aldehydes and acids - Romao, M. J.; Cunha, C. A.; Brondino, C. D.; Moura, J. J. (Met Ions Biol Syst, 2002)
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0161-5149 (Print)0161-5149 (Linking)Journal ArticleResearch Support, Non-U.S. Gov'tReview
Keywords: Acids; Aldehyde Oxidoreductases/chemistry/*metabolism; Aldehydes/*metabolism; Animals; Models, Molecular; Molybdenum/*metabolism; Xanthine Oxidase/chemistry/*metabolism

Molybdenum enzymes in reactions involving aldehydes and acids - Romao, M. J.; Cunha, C. A.; Brondino, C. D.; Moura, J. J. G. (Molybdenum and Tungsten: Their Roles in Biological Processes, 2002)
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Times Cited: 8 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 8

A new approach to the initial value problem - Manuel Ortigueira (5th Portuguese Conference on Automatic Control, 2002)
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O projecto OCTOPUS: O módulo Reconhecedor de Zonas Oxidadas– - Oliveira, R.; Pita, H.; Coito, F.; Steiger-Garção, A. (Jornadas de Engenharia de Telecomunicações e Computadores-ISEL Lisboa, Portugal, 2002)
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Photochromic properties of 3-methyl-substituted flavylium salts - Roque, A.; Lodeiro, C.; Pina, F; Maestri, M; Ballardini, R; Balzani, V. (European Journal of Organic Chemistry, 2002)
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Times Cited: 18 Pina, Fernando/C-8161-2011; Lodeiro, Carlos/B-4793-2013; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, C4O/M-5599-2013; REQUIMTE, UCIBIO/N-9846-2013; REQUIMTE, LAQV/N-9835-2013 Pina, Fernando/0000-0001-8529-6848; Lodeiro, Carlos/0000-0001-5582-5446; 19

Preparation and characterization of new room temperature ionic liquids - Branco, L. C.; Rosa, J. N.; Ramos, J. J. M.; C. A. M. Afonso (Chemistry-a European Journal, 2002)
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Times Cited: 356 Branco, Luis/C-6426-2013; Rosa, Joao/I-6129-2013; REQUIMTE, PHOTO/M-4229-2013; Mateus Afonso, Carlos Alberto/M-7833-2013; REQUIMTE, LAQV/N-9835-2013; iMed.ULisboa, iMed.ULisboa/C-6292-2014; iMed.ULisboa, BioOrgChem /B-4209-2014 Branco, Luis/0000-0003-2520-1151; Rosa, Joao/0000-0001-5165-8820; Mateus Afonso, Carlos Alberto/0000-0002-7284-5948; 374

Proton transfer in anthocyanins and related flavylium salts. Determination of ground-state rate constants with nanosecond laser flash photolysis - Macanita, AL; Moreira, PF; J. C. Lima; Quina, FH; Yihwa, C; Vautier-Giongo, C (Journal of Physical Chemistry a, 2002)
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Rotational friction in AOT microemulsions: Relevance of hydrodynamic and dielectric contributions to microviscosities probed by fluorescent bis 4-(dimethylamino)phenyl squaraine - Laia, C. A. T.; Costa, S. M. B. (Langmuir, 2002)
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Times Cited: 35 Laia, Cesar/C-8651-2011; Costa, Silvia/H-7766-2012; REQUIMTE, PHOTO/M-4229-2013; Costa, Silvia/M-8085-2013; REQUIMTE, LAQV/N-9835-2013 Laia, Cesar/0000-0001-6410-6072; Costa, Silvia/0000-0002-6978-8903; 35

Stoichiometric production of hydrogen peroxide and parallel formation of ferric multimers through decay of the diferric-peroxo complex, the first detectable intermediate in ferritin mineralization - Jameson, GNL; Jin, W; Krebs, C.; Perreira, AS; Tavares, P; Liu, XF; Theil, EC; Huynh, B. H. (Biochemistry, 2002)
The catalytic step that initiates formation of the ferric oxy-hydroxide mineral core in the central cavity of H-type ferritin involves rapid oxidation of ferrous ion by molecular oxygen (ferroxidase reaction) at a binuclear site (ferroxidase site) found in each of the 24 subunits. Previous investigators have shown that the first detectable reaction intermediate of the ferroxidase reaction is a diferric-peroxo intermediate, F-peroxo, formed within 25 ms, which then leads to the release of H2O2 and formation of ferric mineral precursors. The stoichiometric relationship between F-peroxo, H2O2, and ferric mineral precursors, crucial to defining the reaction pathway and mechanism, has now been determined. To this end, a horseradish peroxidase-catalyzed spectrophotometric method was used as an assay for H2O2. By rapidly mixing apo M ferritin from frog, Fe2+, and O-2 and allowing the reaction to proceed for 70 ms when F-peroxo has reached its maximum accumulation, followed by spraying the reaction mixture into the H2O2 assay solution, we were able to quantitatively determine the amount of H2O2 produced during the decay of F-peroxo. The correlation between the amount of H2O2 released with the amount of F-peroxo accumulated at 70 ms determined by Mossbauer spectroscopy showed that F-peroxo decays into H2O2 with a stoichiometry of 1 F-peroxo:H2O2. When the decay of F-peroxo was monitored by rapid freeze-quench Mossbauer spectroscopy, multiple diferric mu-oxo/mu-hydroxo complexes and small polynuclear ferric clusters were found to form at rate constants identical to the decay rate of F-peroxo. This observed parallel formation of multiple products (H2O2, diferric complexes, and small polynuclear clusters) from the decay of a single precursor (F-peroxo) provides useful mechanistic insights into ferritin mineralization and demonstrates a flexible ferroxidase site.
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Studies on the selective transport of organic compounds by using ionic liquids as novel supported liquid membranes - Branco, L. C.; Crespo, J. G.; C. A. M. Afonso (Chemistry-a European Journal, 2002)
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Times Cited: 94 Branco, Luis/C-6426-2013; Crespo, Joao/D-8719-2013; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, CENG/M-4555-2013; Mateus Afonso, Carlos Alberto/M-7833-2013; REQUIMTE, LAQV/N-9835-2013; iMed.ULisboa, iMed.ULisboa/C-6292-2014; iMed.ULisboa, BioOrgChem /B-4209-2014 Branco, Luis/0000-0003-2520-1151; Mateus Afonso, Carlos Alberto/0000-0002-7284-5948; 99

Superoxide reductase activities of neelaredoxin and desulfoferrodoxin metalloproteins - Rusnak, F.; Ascenso, C.; Moura, I.; Moura, J. J. (Methods Enzymol, 2002)
Superoxide reductases have now been well characterized from several organisms. Unique biochemical features include the ability of the reduced enzyme to react with O2- but not dioxygen (reduced SORs are stable in an aerobic atmosphere for hours). Future biochemical assays that measure the reaction of SOR with O2- should take into account the difficulties of assaying O2- directly and the myriad of redox reactions that can take place between components in the assay, for example, direct electron transfer between cytochrome c and Dfx. Future prospects include further delineation of the reaction mechanisms, characterization of the putative (hydro)peroxo intermediate, and studies that uncover the components between reduced pyridine nucleotides and SOR in the metabolic pathway responsible for O2- detoxification.
0076-6879 (Print)0076-6879 (Linking)Journal ArticleReview
Keywords: *Iron-Binding Proteins; Amino Acid Sequence; Bacterial Proteins; Carrier Proteins/chemistry/genetics/*metabolism; Catalytic Domain; Cloning, Molecular; Desulfovibrio/enzymology/genetics; Electron Spin Resonance Spectroscopy; Ferredoxins/chemistry/genetics/*metabolism; Gene Expression; Molecular Sequence Data; Oxidation-Reduction; Oxidoreductases/chemistry/genetics/*metabolism; Recombinant Proteins/genetics/isolation & purification; Sequence Homology, Amino Acid; Spectrophotometry; Spectroscopy, Mossbauer; Superoxide Dismutase/metabolism; Superoxides/metabolism; Treponema pallidum/enzymology/genetics

Switching from intramolecular energy transfer to intramolecular electron transfer by the action of pH and Zn2+ co-ordination (vol 353, pg 63, 2002) - Pina, F (Chemical Physics Letters, 2002)
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Times Cited: 0 Pina, Fernando/C-8161-2011; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Pina, Fernando/0000-0001-8529-6848; 0

Thermal isomerization of a symmetrical carbocyanine molecule: charge transfer aspects - Laia, C. A. T.; Costa, S. M. B. (Chemical Physics Letters, 2002)
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Times Cited: 1 Laia, Cesar/C-8651-2011; Costa, Silvia/H-7766-2012; REQUIMTE, PHOTO/M-4229-2013; Costa, Silvia/M-8085-2013; REQUIMTE, LAQV/N-9835-2013 Laia, Cesar/0000-0001-6410-6072; Costa, Silvia/0000-0002-6978-8903; 1

{Genderforschung und Technikentwicklung [Gender Research and Technology Development: introduction to the main topics]} - Krings, Bettina (University Library of Munich, Germany, 2002)
Since the UN World Conference on Women in 1995 the discussion on gender has been extremely controverse. This issue of the Journal on Technology Assessment (from ITAS) presents and discuss the topic of gender research, and the relation between work and life and the changes that are occuring in the spheres of working lifes and daily lifes. Noch im Umfeld der 4. UN-Weltfrauenkonferenz in Peking 1995 wurde der Begriff gender extrem kontrovers diskutiert. Die Zeitschrift für Technikfolgenabschätzung des ITAS greift mit diesem Schwerpunkt erstmals das Thema „Genderforschung“ auf. Die Verknüpfung der beiden Themengebiete geht auf das Projekt: Wandel der Lebens- und Arbeitsbedingungen im Multimediabereich aus der Genderperspektive zurück, das zurzeit von ITAS im Auftrag des Sozialministerium Baden-Württemberg durchgeführt wird.
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Keywords: gender; Technology Assessment; Work

{Impactos sociais do desinvestimento [Social Impacts of divestment]} - António Moniz; Gomes, Cláudia (University Library of Munich, Germany, 2002)
The resulting economic integration of industrial processes and manufacturing internationalisation lead several authors to argue that world economy is globalised. In this context, the approach to the divestment concept without an social and económical context, does not show a group of associated practices and representations. Choices and options are motivated by exogenous forces that pushes companies to determine strategies that stop capital investment on new equipment goods, or on other imaterial goods. This type of strategy is designated by "divestment". The social level of consequencies are not due to the closing down or de-localization of production units that are divesting, but can be materialised of efects that are irreversible. This means unemployment, de-skilling, labour precarization and even emergence of new forms of social exclusion in former industrialised regions.
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Keywords: di-vestment; globalization; labour markets; strategical management

A cooperação entre equipas de trabalho em empresas em rede: vantagens para o desenvolvimento regional [Workteam Co-operation in Networked Companies: regional development advantages] - António Moniz (University Library of Munich, Germany, 2001)
Working teams in enterprise environment are considered as the most advanced forms of work organisation. This means the forms that can improve productivity quality of working life. Nevertheless, it prevail a slow development and dissemination of these advanced organisational forms in European companies. The reason for that lays in a complex linkage factors from social values to the economical pressures. But other factors are also related to the national systems of education training, to the different systems of industrial relations and technology policy.
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Keywords: enterprise networks; Europe; innovation; labour; productivity; work organization

{A cooperação entre equipas de trabalho em empresas em rede: vantagens para o desenvolvimento regional [Workteam Co-operation in Networked Companies: regional development advantages]} - António Moniz (University Library of Munich, Germany, 2001)
Working teams in enterprise environment are considered as the most advanced forms of work organisation. This means the forms that can improve productivity quality of working life. Nevertheless, it prevail a slow development and dissemination of these advanced organisational forms in European companies. The reason for that lays in a complex linkage factors from social values to the economical pressures. But other factors are also related to the national systems of education training, to the different systems of industrial relations and technology policy.
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Keywords: enterprise networks; Europe; innovation; labour; productivity; work organization

Structure refinement of the aldehyde oxidoreductase from Desulfovibrio gigas (MOP) at 1.28 A - Rebelo, J. M.; Dias, J. M.; Huber, R.; Moura, J. J.; Romao, M. J. (J Biol Inorg Chem, 2001)
The sulfate-reducing bacterium aldehyde oxidoreductase from Desulfovibrio gigas (MOP) is a member of the xanthine oxidase family of enzymes. It has 907 residues on a single polypeptide chain, a molybdopterin cytosine dinucleotide (MCD) cofactor and two [2Fe-2S] iron-sulfur clusters. Synchrotron data to almost atomic resolution were collected for improved cryo-cooled crystals of this enzyme in the oxidized form. The cell constants of a=b=141.78 A and c=160.87 A are about 2% shorter than those of room temperature data, yielding 233,755 unique reflections in space group P6(1)22, at 1.28 A resolution. Throughout the entire refinement the full gradient least-squares method was used, leading to a final R factor of 14.5 and Rfree factor of 19.3 (4sigma cut-off) with "riding" H-atoms at their calculated positions. The model contains 8146 non-hydrogen atoms described by anisotropic displacement parameters with an observations/parameters ratio of 4.4. It includes alternate conformations for 17 amino acid residues. At 1.28 A resolution, three Cl- and two Mg2+ ions from the crystallization solution were clearly identified. With the exception of one Cl- which is buried and 8 A distant from the Mo atom, the other ions are close to the molecular surface and may contribute to crystal packing. The overall structure has not changed in comparison to the lower resolution model apart from local corrections that included some loop adjustments and alternate side-chain conformations. Based on the estimated errors of bond distances obtained by blocked least-squares matrix inversion, a more detailed analysis of the three redox centres was possible. For the MCD cofactor, the resulting geometric parameters confirmed its reduction state as a tetrahydropterin. At the Mo centre, estimated corrections calculated for the Fourier ripples artefact are very small when compared to the experimental associated errors, supporting the suggestion that the fifth ligand is a water molecule rather than a hydroxide. Concerning the two iron-sulfur centres, asymmetry in the Fe-S distances as well as differences in the pattern of NH.S hydrogen-bonding interactions was observed, which influences the electron distribution upon reduction and causes non-equivalence of the individual Fe atoms in each cluster.
0949-8257 (Print)0949-8257 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Aldehyde Oxidoreductases/*chemistry; Bacterial Proteins/chemistry; Chlorides/chemistry; Crystallization; Crystallography, X-Ray; Desulfovibrio/chemistry/*enzymology; Iron-Sulfur Proteins/*chemistry; Least-Squares Analysis; Magnesium/chemistry; Models, Molecular; Molybdenum/*chemistry; Protein Conformation; Pterins/chemistry

Characterization of recombinant Desulfovibrio gigas ferredoxin - Rodrigues, P.; Graca, F.; Macedo, A. L.; Moura, I.; Moura, J. J. (Biochem Biophys Res Commun, 2001)
Dg ferredoxin gene was cloned using the polymerase chain reaction (PCR), inserted into vector pT7-7, and overexpressed in Escherichia coli (E. coli) grown in aerobic media. The recombinant protein is a dimer and contains a [3Fe-4S] cluster per monomer. EPR and (1)H NMR data of recombinant and wild-type protein are compared.
0006-291X (Print)0006-291X (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Cloning, Molecular; Desulfovibrio/*chemistry; Dimerization; Electron Spin Resonance Spectroscopy; Electrophoresis, Polyacrylamide Gel; Escherichia coli/metabolism; Ferredoxins/*chemistry/genetics; Iron/metabolism; Magnetic Resonance Spectroscopy; Oxygen/metabolism; Polymerase Chain Reaction; Recombinant Proteins/*chemistry/metabolism; Spectrophotometry; Sulfur/metabolism; Temperature; Ultraviolet Rays

Protein effects on the electronic structure of the [Fe4S4]2+ cluster in ferredoxin and HiPIP - Glaser, T.; Bertini, I.; Moura, J. J.; Hedman, B.; Hodgson, K. O.; Solomon, E. I. (J Am Chem Soc, 2001)
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0002-7863 (Print)0002-7863 (Linking)Journal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, Non-P.H.S.Research Support, U.S. Gov't, P.H.S.
Keywords: *Photosynthetic Reaction Center Complex Proteins; Bacterial Proteins; Chemistry, Physical; Ferredoxins/*chemistry; Iron-Sulfur Proteins/*chemistry; Molecular Structure; Physicochemical Phenomena; Spectrum Analysis/methods; X-Rays

Substitution of murine ferrochelatase glutamate-287 with glutamine or alanine leads to porphyrin substrate-bound variants - Franco, R.; Pereira, A. S.; Tavares, P; Mangravita, A.; Barber, M. J.; Moura, I.; Ferreira, G. C. (Biochemical Journal, 2001)
Ferrochelatase (EC 4.99.1.1) is the terminal enzyme of the haem biosynthetic pathway and catalyses iron chelation into the protoporphyrin IX ring. Glutamate-287 (E287) of murine mature ferrochelatase is a conserved residue in all known sequences of ferrochelatase, is present at the active site of the enzyme, as inferred from the Bacillus subtilis ferrochelatase three-dimensional structure, and is critical for enzyme activity. Substitution of E287 with either glutamine (Q) or alanine (A) yielded variants with lower enzymic activity than that of the wild-type ferrochelatase and with different absorption spectra from the wild-type enzyme. In contrast to the wild-type enzyme, the absorption spectra of the variants indicate that these enzymes, as purified, contain protoporphyrin IX. Identification and quantification of the porphyrin bound to the E287-directed variants indicate that approx. 80% of the total porphyrin corresponds to protoporphyrin IX. Significantly, rapid stopped-flow experiments of the E287A and E287Q Variants demonstrate that reaction with Zn2+ results in the formation of bound Zn-protoporphyrin IX, indicating that the endogenously bound protoporphyrin IX can be used as a substrate. Taken together, these findings suggest that the structural strain imposed by ferrochelatase on the porphyrin substrate as a critical step in the enzyme catalytic mechanism is also accomplished by the E287A and E287Q variants, but without the release of the product. Thus E287 in murine ferrochelatase appears to be critical For the catalytic process by controlling the release of the product.
Times Cited: 14Part 1

Mossbauer characterization of the iron-sulfur clusters in Desulfovibrio vulgaris hydrogenase - Pereira, A. S.; Tavares, P; Moura, I.; Moura, J. J.; Huynh, B. H. (J Am Chem Soc, 2001)
The periplasmic hydrogenase of Desulfovibrio vulgaris (Hildenbourough) is an all Fe-containing hydrogenase. It contains two ferredoxin type [4Fe-4S] clusters, termed the F clusters, and a catalytic H cluster. Recent X-ray crystallographic studies on two Fe hydrogenases revealed that the H cluster is composed of two sub-clusters, a [4Fe-4S] cluster ([4Fe-4S](H)) and a binuclear Fe cluster ([2Fe](H)), bridged by a cysteine sulfur. The aerobically purified D. vulgaris hydrogenase is stable in air. It is inactive and requires reductive activation. Upon reduction, the enzyme becomes sensitive to O(2), indicating that the reductive activation process is irreversible. Previous EPR investigations showed that upon reoxidation (under argon) the H cluster exhibits a rhombic EPR signal that is not seen in the as-purified enzyme, suggesting a conformational change in association with the reductive activation. For the purpose of gaining more information on the electronic properties of this unique H cluster and to understand further the reductive activation process, variable-temperature and variable-field Mossbauer spectroscopy has been used to characterize the Fe-S clusters in D. vulgaris hydrogenase poised at different redox states generated during a reductive titration, and in the CO-reacted enzyme. The data were successfully decomposed into spectral components corresponding to the F and H clusters, and characteristic parameters describing the electronic and magnetic properties of the F and H clusters were obtained. Consistent with the X-ray crystallographic results, the spectra of the H cluster can be understood as originating from an exchange coupled [4Fe-4S]-[2Fe] system. In particular, detailed analysis of the data reveals that the reductive activation begins with reduction of the [4Fe-4S](H) cluster from the 2+ to the 1+ state, followed by transfer of the reducing equivalent from the [4Fe-4S](H) subcluster to the binuclear [2Fe](H) subcluster. The results also reveal that binding of exogenous CO to the H cluster affects significantly the exchange coupling between the [4Fe-4S](H) and the [2Fe](H) subclusters. Implication of such a CO binding effect is discussed.
0002-7863 (Print)0002-7863 (Linking)Journal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.
Keywords: Desulfovibrio vulgaris/*enzymology; Hydrogenase/*chemistry/isolation & purification/metabolism; Iron-Sulfur Proteins/*chemistry/metabolism; Molecular Structure; Oxidation-Reduction; Spectroscopy, Mossbauer

Effects of bilirubin molecular species on membrane dynamic properties of human erythrocyte membranes: a spin label electron paramagnetic resonance spectroscopy study - Brito, M. A.; Brondino, C. D.; Moura, J. J.; Brites, D. (Arch Biochem Biophys, 2001)
Unconjugated bilirubin is a neurotoxic pigment that interacts with membrane lipids. In this study we used electron paramagnetic resonance and the spin labels 5-, 7-, 12-, and 16-doxyl-stearic acid (DSA) to evaluate the depth of the hydrocarbon chain at which interaction of bilirubin preferentially occurs. In addition, we used different pH values to determine the molecular species involved. Resealed right-side-out ghosts were incubated (1-60 min) with bilirubin (3.4-42.8 microM) at pH 7.0, 7.4, and 8.0. Alterations of membrane dynamic properties were maximum after 15 min of incubation with 8.6 microM bilirubin at pH 7.4 and were accompanied by a significant release of phospholipids. Interestingly, concentrations of bilirubin up to 42.8 microM and longer incubations resulted in the elution of cholesterol and further increased that of phospholipids while inducing less structural alterations. Variation of the pH values from 8.0 to 7.4 and 7.0, under conditions of maximum perturbation, led to a change from an increased to a diminished polarity sensed by 5-DSA. Conversely, a progressive enhancement in fluidity was reported by 7-DSA, followed by 12- and 16-DSA. These results indicate that bilirubin while enhancing membrane lipid order at C-5 simultaneously has disordering effects at C-7. Furthermore, recovery of membrane dynamics after 15 min of bilirubin exposure along with the release of lipids is compatible with a membrane adaptive response to the insult. In addition, our data provide evidence that uncharged diacid is the species primarily interacting with the membrane as perturbation is favored by acidosis, a condition frequently associated with hyperbilirubinemia in premature and severely ill infants.
0003-9861 (Print)0003-9861 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Bilirubin/*pharmacology; Cholesterol/analysis; Electron Spin Resonance Spectroscopy; Erythrocyte Membrane/*drug effects; Humans; Membrane Fluidity/drug effects; Phospholipids/chemistry; Spin Labels; Stearic Acids/chemistry

A análise prospectiva como ferramenta de pol{\'ıtica de inovação: uma perspectiva socioeconómica [Foresight analysis as an innovation policy tool: a socio-economical approach] - António Moniz; Godinho, Manuel M. (University Library of Munich, Germany, 2001)
At the last National Conference of Industrial Sociology in Portugal (in March 1999) was presented a paper on the topic of “foresight as a technology and employment policy instrument” (A.B. Moniz) where there was a reference to the need of development of this kind of tools and instruments. This need is clear when one should neutralize the negative effects of such policies, and could support the positive influences, especially with the main aims of improvement of quality of working life. Thus, is today possible to make a first balance of the application in Portugal of a type of foresight analysis exercise: the Delphi method. After some of the uses of this method in Portugal on the fisheries socio-economical system and on the relation between information society and employment, we present in this paper a first assessment of such experiments and present some recommendations for future uses of this foresight technique. We conclude that they must be applied within the context of socio-economical perspectives, and not only the technological ones, although they should include that dimension.
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Keywords: employment policy; foresight; Portugal; socio-economical s; Technology Assessment; technology policy

Novos Modelos de Produção na Indústria Automóvel Algumas Interrogações [New models of production in automotive industry: some questions] - António Moniz; Machado, Tiago (University Library of Munich, Germany, 2001)
Taking into account the global trends towards vertical de-integration and functional integration, the WorTiS project is expected to be able to determine to what extent the Portuguese automobile industry is experiencing far-reaching changes as far as innovative (post-fordist) work systems are concerned. One of the objectives is to reach wider conclusions regarding the sector under analysis within a new multi-disciplinary approach, in connection with other research networks (namely, GERPISA and IMVP-MIT). It will recover relevant information in automobile companies located in Portugal (like, Toyota, Citröen, FIAT, Renault, Ford, VW, UMM), in order to understand how concrete practices have being developed in time, and update the scientific knowledge with the development of new case studies (Mitsubishi, AutoEuropa, Opel-GM, and other sub-contracting firms). is intended to present a new fieldwork methodology in order to analyse the variety, and contradictory character, of changes in work practices. The analysis would focus on the cases of automotive firms that operated, and are still operating, in Portugal. The effectiveness of such a tool will last far beyond the project itself.
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Keywords: automotive industry; production models; sociology; working systems

Production of brown tyrosine pigments by the yeast Yarrowia lipolytica - Carreira, A; Ferreira, LM; Loureiro, V (JOURNAL OF APPLIED MICROBIOLOGY, 2001)
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{A análise prospectiva como ferramenta de política de inovação: uma perspectiva socioeconómica [Foresight analysis as an innovation policy tool: a socio-economical approach]} - António Moniz; Godinho, Manuel M. (University Library of Munich, Germany, 2001)
At the last National Conference of Industrial Sociology in Portugal (in March 1999) was presented a paper on the topic of “foresight as a technology and employment policy instrument” (A.B. Moniz) where there was a reference to the need of development of this kind of tools and instruments. This need is clear when one should neutralize the negative effects of such policies, and could support the positive influences, especially with the main aims of improvement of quality of working life. Thus, is today possible to make a first balance of the application in Portugal of a type of foresight analysis exercise: the Delphi method. After some of the uses of this method in Portugal on the fisheries socio-economical system and on the relation between information society and employment, we present in this paper a first assessment of such experiments and present some recommendations for future uses of this foresight technique. We conclude that they must be applied within the context of socio-economical perspectives, and not only the technological ones, although they should include that dimension.
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Keywords: employment policy; foresight; Portugal; socio-economical s; Technology Assessment; technology policy

{Novos Modelos de Produção na Indústria Automóvel Algumas Interrogações [New models of production in automotive industry: some questions]} - António Moniz; Machado, Tiago (University Library of Munich, Germany, 2001)
Taking into account the global trends towards vertical de-integration and functional integration, the WorTiS project is expected to be able to determine to what extent the Portuguese automobile industry is experiencing far-reaching changes as far as innovative (post-fordist) work systems are concerned. One of the objectives is to reach wider conclusions regarding the sector under analysis within a new multi-disciplinary approach, in connection with other research networks (namely, GERPISA and IMVP-MIT). It will recover relevant information in automobile companies located in Portugal (like, Toyota, Citröen, FIAT, Renault, Ford, VW, UMM), in order to understand how concrete practices have being developed in time, and update the scientific knowledge with the development of new case studies (Mitsubishi, AutoEuropa, Opel-GM, and other sub-contracting firms). is intended to present a new fieldwork methodology in order to analyse the variety, and contradictory character, of changes in work practices. The analysis would focus on the cases of automotive firms that operated, and are still operating, in Portugal. The effectiveness of such a tool will last far beyond the project itself.
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Keywords: automotive industry; production models; sociology; working systems

Calcium-dependent conformation of a heme and fingerprint peptide of the diheme cytochrome c peroxidase from Paracoccus pantotrophus - Pauleta, S. R.; Lu, Y.; Goodhew, C. F.; Moura, I.; Pettigrew, G. W.; Shelnutt, J. A. (Biochemistry, 2001)
The structural changes in the heme macrocycle and substituents caused by binding of Ca2+ to the diheme cytochrome c peroxidase from Paracoccus pantotrophus were clarified by resonance Raman spectroscopy of the inactive fully oxidized form of the enzyme. The changes in the macrocycle vibrational modes are consistent with a Ca2+-dependent increase in the out-of-plane distortion of the low-potential heme, the proposed peroxidatic heme. Most of the increase in out-of-plane distortion occurs when the high-affinity site I is occupied, but a small further increase in distortion occurs when site II is also occupied by Ca2+ or Mg2+. This increase in the heme distortion explains the red shift in the Soret absorption band that occurs upon Ca2+ binding. Changes also occur in the low-frequency substituent modes of the heme, indicating that a structural change in the covalently attached fingerprint pentapeptide of the LP heme occurs upon Ca2+ binding to site I. These structural changes may lead to loss of the sixth ligand at the peroxidatic heme in the semireduced form of the enzyme and activation.
Times Cited: 8

El desarrollo socio-cultural de la sociedad de la información: Competencia o cooperación? [Socio-cultural development of information society: competition or co-operation?] - Krings, Bettina; António Moniz (University Library of Munich, Germany, 2001)
The results presented in this article are related to the field of investigation defined inside the European TSER project on "Information Society, Work and Generation of New Forms of Social Exclusion " (SOWING). The thematic center of the project is connected with the question of the relations that exist between the introduction of information technologies (IT) and the changes in the waged labour as well as in the organisation of work systems. The introduction of these technologies transformed specially the activities on services as well as certain branches, where great changes of the organization of work for the next future assume, which are discussed within the margin of the debate on future forms of work. The tendencies within the occupational structures indicate an individualisation of the insecurities and an opening of the occupational carreers. For these reasons, the development of the occupational carreers is more and more bound to the biographical planning. The institutionalised occupational model is changing very strongly, which in the long term will take to social and cultural changes, not only in the professional life, but also in the private life.
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Keywords: Information Society; information technology; occupations; Work

Estratégias de construção da Sociedade da Informação [Strategies of construction of Information Society] - António Moniz (University Library of Munich, Germany, 2001)
The most significative text of the European Commission on the construction of a strategy for an Information Society is, undoubtly the document on “Job Opportunities in the Information Society”. The major dimension of the change process was not anymore the technological evolution one. Information society should integrate the social-economical dimensions with industries that use information and communication products and services, beside those of so-called “Information Society industries”. Is not only this sector that is growing in terms of employment, but also the other sectors can be also important ICT users. Employment in the information society leads the employment growth in EU. In this study are analysed the different national programs and strategies, and with particular relevance the Portuguese case.
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Keywords: employment; European Union; Information Society; Portugal

Technological practices in the European auto industry: Exploring cases from Belgium, Germany and Portugal - António Moniz; Krings, Bettina; Van Hootegem, Geert; Huys, Rik (University Library of Munich, Germany, 2001)
The relation between work organisation and technological practices in auto industry is analysed in this article. The concept of “technological practice” in this sector is used to describe the specific ways of embedding information and communication technology applications into the organizational forms and cultural patterns. This concept was developed with the Sowing project (TSER, DG XII) and that approach included either the shop floor co-operation up to the regionally based networks of companies and supporting institutions. The authors studied different sectors in the automotive firms of different European countries (Germany, Belgium and Portugal): shopfloor and production lines, design and management and the local inter-relationships. It was underlined some evidencies of the different alternatives in terms of technological practices for the same sector. Much of the litterature try to disseminate an idea of a single (and optimum) organisational model for the same type of product. And here, even with the same type of technology, and of product (medium-high range), one can find different models, different cultures, different ways of organising the industrial structure (firms, regional institutions, R&D centres) in the same sector (auto industry).
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Keywords: automobile sector; Information and Communication Technologies; technological practice; work organisa

{El desarrollo socio-cultural de la sociedad de la información: Competencia o cooperación? [Socio-cultural development of information society: competition or co-operation?]} - Krings, Bettina; António Moniz (University Library of Munich, Germany, 2001)
The results presented in this article are related to the field of investigation defined inside the European TSER project on "Information Society, Work and Generation of New Forms of Social Exclusion " (SOWING). The thematic center of the project is connected with the question of the relations that exist between the introduction of information technologies (IT) and the changes in the waged labour as well as in the organisation of work systems. The introduction of these technologies transformed specially the activities on services as well as certain branches, where great changes of the organization of work for the next future assume, which are discussed within the margin of the debate on future forms of work. The tendencies within the occupational structures indicate an individualisation of the insecurities and an opening of the occupational carreers. For these reasons, the development of the occupational carreers is more and more bound to the biographical planning. The institutionalised occupational model is changing very strongly, which in the long term will take to social and cultural changes, not only in the professional life, but also in the private life.
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Keywords: Information Society; information technology; occupations; Work

{El desarrollo socio-cultural de la sociedad de la información: Competencia o cooperación? [Socio-cultural development of information society: competition or co-operation?]} - Krings, Bettina; António Moniz (University Library of Munich, Germany, 2001)
The results presented in this article are related to the field of investigation defined inside the European TSER project on "Information Society, Work and Generation of New Forms of Social Exclusion " (SOWING). The thematic center of the project is connected with the question of the relations that exist between the introduction of information technologies (IT) and the changes in the waged labour as well as in the organisation of work systems. The introduction of these technologies transformed specially the activities on services as well as certain branches, where great changes of the organization of work for the next future assume, which are discussed within the margin of the debate on future forms of work. The tendencies within the occupational structures indicate an individualisation of the insecurities and an opening of the occupational carreers. For these reasons, the development of the occupational carreers is more and more bound to the biographical planning. The institutionalised occupational model is changing very strongly, which in the long term will take to social and cultural changes, not only in the professional life, but also in the private life.
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Keywords: Information Society; information technology; occupations; Work

{Estratégias de construção da Sociedade da Informação [Strategies of construction of Information Society]} - António Moniz (University Library of Munich, Germany, 2001)
The most significative text of the European Commission on the construction of a strategy for an Information Society is, undoubtly the document on “Job Opportunities in the Information Society”. The major dimension of the change process was not anymore the technological evolution one. Information society should integrate the social-economical dimensions with industries that use information and communication products and services, beside those of so-called “Information Society industries”. Is not only this sector that is growing in terms of employment, but also the other sectors can be also important ICT users. Employment in the information society leads the employment growth in EU. In this study are analysed the different national programs and strategies, and with particular relevance the Portuguese case.
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Keywords: employment; European Union; Information Society; Portugal

{Technological practices in the European auto industry: Exploring cases from Belgium, Germany and Portugal} - António Moniz; Krings, Bettina; Van Hootegem, Geert; Huys, Rik (University Library of Munich, Germany, 2001)
The relation between work organisation and technological practices in auto industry is analysed in this article. The concept of “technological practice” in this sector is used to describe the specific ways of embedding information and communication technology applications into the organizational forms and cultural patterns. This concept was developed with the Sowing project (TSER, DG XII) and that approach included either the shop floor co-operation up to the regionally based networks of companies and supporting institutions. The authors studied different sectors in the automotive firms of different European countries (Germany, Belgium and Portugal): shopfloor and production lines, design and management and the local inter-relationships. It was underlined some evidencies of the different alternatives in terms of technological practices for the same sector. Much of the litterature try to disseminate an idea of a single (and optimum) organisational model for the same type of product. And here, even with the same type of technology, and of product (medium-high range), one can find different models, different cultures, different ways of organising the industrial structure (firms, regional institutions, R&D centres) in the same sector (auto industry).
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Keywords: automobile sector; Information and Communication Technologies; technological practice; work organisa

Kinetics of inter- and intramolecular electron transfer of Pseudomonas nautica cytochrome cd1 nitrite reductase: regulation of the NO-bound end product - Lopes, H.; Besson, S.; Moura, I.; Moura, J. J. (J Biol Inorg Chem, 2001)
The intermolecular electron transfer kinetics between nitrite reductase (NiR, cytochrome cd1) isolated from Pseudomonas nautica and three cytochromes c isolated from the same strain, as well as the intramolecular electron transfer between NiR heme c and NiR heme d1, were investigated by cyclic voltammetry. All cytochromes (cytochrome c552, cytochrome c553 and cytochrome C553(548)) exhibited well-behaved electrochemistry. The individual diffusion coefficients and mid-point redox potentials were determined. Under the experimental conditions, only cytochrome c552 established a rapid electron transfer with NiR. At acidic pH, the intermolecular electron transfer (cytochrome c(552red)-->NiR heme cox) is a second-order reaction with a rate constant (k2) of 4.1+/-0.1x10(5) M(-1) s(-1) (pH=6.3 and 100 mM NaCl). Under these conditions, the intermolecular reaction represents the rate-limiting step. A minimum estimate of 33 s(-1) could be determined for the first-order rate constant (k1) of the intramolecular electron transfer reaction NiR heme c(red)-->NiR heme d1ox. The pH dependence of k2 values was investigated at pH values ranging from 5.8 to 8.0. When the pH is progressively shifted towards basic values, the rate constant of the intramolecular electron transfer reaction NiR heme c(red)-->NiR heme d1ox decreases gradually to a point where it becomes rate limiting. At pH 8.0 we determined a value of 1.4+/-0.7 s(-1), corresponding to a k2 value of 2.2+/-1.1x10(4) M(-1) s(-1) for the intermolecular step. The physiological relevance of these results is discussed with a particular emphasis on the proposed mechanism of "dead-end product" formation.
0949-8257 (Print)0949-8257 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Catalysis; Cytochromes; Electrochemistry; Electron Transport; Electron Transport Complex IV/chemistry/*metabolism; Kinetics; Nitrite Reductases/chemistry/*metabolism; Pseudomonas/*enzymology

Amyloid beta-peptide disrupts mitochondrial membrane lipid and protein structure: protective role of tauroursodeoxycholate - Rodrigues, C. M.; Sola, S.; Brito, M. A.; Brondino, C. D.; Brites, D.; Moura, J. J. (Biochem Biophys Res Commun, 2001)
Mitochondria have been implicated in the cytotoxicity of amyloid beta-peptide (A beta), which accumulates as senile plaques in the brain of Alzheimer's disease patients. Tauroursodeoxycholate (TUDC) modulates cell death, in part, by preventing mitochondrial membrane perturbation. Using electron paramagnetic resonance spectroscopy analysis of isolated mitochondria, we tested the hypothesis that A beta acts locally in mitochondrial membranes to induce oxidative injury, leading to increased membrane permeability and subsequent release of caspase-activating factors. Further, we intended to determine the role of TUDC at preventing A beta-induced mitochondrial membrane dysfunction. The results demonstrate oxidative injury of mitochondrial membranes during exposure to A beta and reveal profound structural changes, including modified membrane lipid polarity and disrupted protein mobility. Cytochrome c is released from the intermembrane space of mitochondria as a consequence of increased membrane permeability. TUDC, but not cyclosporine A, almost completely abrogated A beta-induced perturbation of mitochondrial membrane structure. We conclude that A beta directly induces cytochrome c release from mitochondria through a mechanism that is accompanied by profound effects on mitochondrial membrane redox status, lipid polarity, and protein order. TUDC can directly suppress A beta-induced disruption of the mitochondrial membrane structure, suggesting a neuroprotective role for this bile salt.
0006-291X (Print)0006-291X (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Amyloid beta-Peptides/*pharmacology; Animals; Cytochrome c Group/drug effects/metabolism; Electron Spin Resonance Spectroscopy; Intracellular Membranes/chemistry/*drug effects/metabolism; Membrane Lipids/chemistry/*metabolism; Membrane Proteins/chemistry/*drug effects/metabolism; Mitochondria, Liver/drug effects; Mitochondria/drug effects; Oxidative Stress; Peptide Fragments/pharmacology; Permeability/drug effects; Rats; Rats, Wistar; Spin Labels; Taurochenodeoxycholic Acid/*pharmacology

DOSY studies of hydrogen bond association: tetramethylsilane as a reference compound for diffusion studies - E. J. Cabrita; Berger, S (Magnetic Resonance in Chemistry, 2001)
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Brown pigments produced by Yarrowia lipolytica result from extracellular accumulation of homogentisic acid - Carreira, A; Ferreira, LM; Loureiro, V (APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2001)
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Calcium in bacterial peroxidases - Pseudomonas stutzeri cytochrome c peroxidase - Timóteo, C G; Tavares, P; Pettigrew, G. W.; Moura, I. (Journal Of Inorganic Biochemistry, 2001)
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Cloning of a novel Mo-Cu containing protein from Desulfovibrio.gigas - Di Rocco, G.; Pereira, A. S.; Bursakov, S. A.; Gavel, O. Y.; Rusnak, F.; Lampreia, J.; Moura, J. J. G.; Moura, I. (Journal Of Inorganic Biochemistry, 2001)
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Developmen of an electrochemical biosensor for nitrite determination - Almeida, G.; Tavares, P; Lampreia, J.; Moura, J. J. G.; Moura, I. (Journal Of Inorganic Biochemistry, 2001)
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Nitrous oxide reductase (N2OR) from Pseudomonas nautica 617 - Cabrito, I.; Pereira, A. S.; Tavares, P; Besson, S.; Brondino, C.; Hoffman, B.; Brown, K.; Tegoni, M.; Cambillau, C.; Moura, J. J. G.; Moura, I. (Journal Of Inorganic Biochemistry, 2001)
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Structure-function studies of cytochrome c peroxidase from ps. nautica - Alves, T.; Besson, S.; Pereira, A. S.; Pettigrew, G. W.; Moura, J. J. G.; Moura, I. (Journal Of Inorganic Biochemistry, 2001)
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Imide-amide rearrangement of cyclic phosphorimidates: A mechanistic study - E. J. Cabrita; C. A. M. Afonso; Santos, AGD (Chemistry-a European Journal, 2001)
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Structural aspects of denitrifying enzymes - Moura, I.; Moura, J. J. (Curr Opin Chem Biol, 2001)
The reduction of nitrate to nitrogen gas via nitrite, nitric oxide and nitrous oxide is the metabolic pathway usually known as denitrification, a key step in the nitrogen cycle. As observed for other elemental cycles, a battery of enzymes are utilized, namely the reductases for nitrate, nitrite, nitric oxide and nitrous oxide, as well as multiple electron donors that interact with these enzymes, in order to carry out the stepwise reactions that involve key intermediates. Because of the importance of this pathway (of parallel importance to the nitrogen-fixation pathway), efforts are underway to understand the structures of the participating enzymes and to uncover mechanistic aspects. Three-dimensional structures have been solved for the majority of these enzymes in the past few years, revealing the architecture of the active metal sites as well as global structural aspects, and possible mechanistic aspects. In addition, the recognition of specific electron-transfer partners raises important questions regarding specific electron-transfer pathways, partner recognition and control of metabolism.
1367-5931 (Print)1367-5931 (Linking)Journal ArticleResearch Support, Non-U.S. Gov'tReview
Keywords: Electron Transport; Enzymes/*chemistry/metabolism; Metalloproteins/chemistry; Models, Molecular; Nitrate Reductases/*chemistry/metabolism; Nitric Oxide/metabolism; Nitrite Reductases/*chemistry/metabolism; Nitrous Oxide/metabolism; Oxidoreductases/*chemistry/metabolism; Protein Structure, Tertiary

Tungsten-containing formate dehydrogenase from Desulfovibrio gigas: metal identification and preliminary structural data by multi-wavelength crystallography - Raaijmakers, H.; Teixeira, S.; Dias, J. M.; Almendra, M. J.; Brondino, C. D.; Moura, I.; Moura, J. J.; Romao, M. J. (J Biol Inorg Chem, 2001)
The tungsten-containing formate dehydrogenase (W-FDH) isolated from Desulfovibrio gigas has been crystallized in space group P2(1), with cell parameters a = 73.8 A, b = 111.3 A, c = 156.6 A and beta = 93.7 degrees. These crystals diffract to beyond 2.0 A on a synchrotron radiation source. W-FDH is a heterodimer (92 kDa and 29 kDa subunits) and two W-FDH molecules are present in the asymmetric unit. Although a molecular replacement solution was found using the periplasmic nitrate reductase as a search model, additional phasing information was needed. A multiple-wavelength anomalous dispersion (MAD) dataset was collected at the W- and Fe-edges, at four different wavelengths. Anomalous and dispersive difference data allowed us to unambiguously identify the metal atoms bound to W-FDH as one W atom with a Se-cysteine ligand as well as one [4Fe-4S] cluster in the 92 kDa subunit, and three additional [4Fe-4S] centers in the smaller 29 kDa subunit. The D. gigas W-FDH was previously characterized based on metal analysis and spectroscopic data. One W atom was predicted to be bound to two molybdopterin guanine dinucleotide (MGD) pterin cofactors and two [4Fe-4S] centers were proposed to be present. The crystallographic data now reported reveal a selenium atom (as a Se-cysteine) coordinating to the W site, as well as two extra [4Fe-4S] clusters not anticipated before. The EPR data were re-evaluated in the light of these new results.
0949-8257 (Print)0949-8257 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: *Tungsten; Crystallography, X-Ray/methods; Desulfovibrio/*enzymology; Electron Spin Resonance Spectroscopy; Formate Dehydrogenases/*chemistry; Iron-Sulfur Proteins/chemistry; Metals/analysis; Models, Molecular; Protein Conformation

Exploring the photocatalytic properties and the long-lifetime chemosensor ability of Cl-2 Ru(Bpy)(2)L (L=2,5,8,11,14-pentaaza 15 )-2,2 '-bipyridilophane) - Lodeiro, C.; Pina, F; A. J. Parola; Bencini, A; Bianchi, A; Bazzicalupi, C; Ciattini, S.; Giorgi, C; Masotti, A.; Valtancoli, B; de Melo, JS (Inorganic Chemistry, 2001)
In this work a new water-soluble long-lifetime chemosensor, containing a polyamine unit connected to a complexed Ru(II) metal center, is described. Its crystal structure has been characterized by X-ray analysis. The polyamine macrocyclic unit is capable of anchoring cationic or anionic substrates, according to its protonation state. Examples of electron transfer involving the ruthenium complex core and the bound substrate are presented. The photocatalytic ability of such a system is illustrated by the oxidation of iodide to iodine promoted by light absorption at 436 nm.
ISI Document Delivery No.: 502NHTimes Cited: 16 Cited Reference Count: 53 Lodeiro, C Pina, F Parola, AJ Bencini, A Bianchi, A Bazzicalupi, C Ciattini, S Giorgi, C Masotti, A Valtancoli, B de Melo, JS Masotti, Andrea/D-5306-2009; Seixas de Melo, Sergio/A-5388-2008; Pina, Fernando/C-8161-2011; Lodeiro, Carlos/B-4793-2013; Parola, A. Jorge/F-4048-2010 Masotti, Andrea/0000-0002-2511-5088; Seixas de Melo, Sergio/0000-0001-9708-5079; Lodeiro, Carlos/0000-0001-5582-5446; Parola, A. Jorge/0000-0002-1333-9076 Amer chemical soc Washington

Metal-metal interaction in polynuclear complexes with cyanide bridges: synthesis, characterisation, and theoretical studies - Calhorda, M. J.; Drew, M. G. B.; Felix, V.; Fonseca, L. P.; Gamelas, C. A.; Godinho, Ssmc; Goncalves, I. S.; Hunstock, E.; Lopes, J. P.; A. J. Parola; Pina, F; Romao, C. C.; et al. (Journal of Organometallic Chemistry, 2001)
The reaction of the cyanide anion [M(CO)(5)CN](-) (M = Cr or Mo) with metallocenes of Groups 4 and 6 produced polynuclear complexes of the type [CpCp 'M(CO){-NC-M ' (CO)(5)}]BF4 (M = M0, W; M ' = Mo, Cr, Cp '= Cp, Ind), Cp2TiCl{-NC-Mo(CO)(5)} and Cp2Ti{-NC-Mo(CO)(5)}(2). These complexes were characterised by H-1-, C-13- and Mo-95-NMR, IR and UV-vis spectroscopies, elemental analysis and examined by cyclic voltammetry. These methods show that the [M(CO)(5)CN]- ligands shift the electron density towards the metallocene centres. The complex [Cp2W(CO){-NC-Mo(CO)(5)}](+) is additionally examined by single crystal X-ray structure determination. The Density Functional Theory (DFT) calculations with the ADF program were performed on selected compounds to understand the nature of the redox processes taking place. Compared with a nitrile, the coordination of a [M(CO)-,CN]- fragment to the metallocene moiety does not significantly change the geometrical features. but leads to the stabilisation of the HOMO of the latter. with all the oxidation processes occurring in the pentacarbonyl moiety of the binuclear species. Time-dependent DFT calculations were used to identify the band appearing in the visible spectrum of Cp2TiCl{-NC-Mo(CO)(5)} as a Mo to Ti charge transfer. (C) 2001 Elsevier Science BN. All rights reserved.
ISI Document Delivery No.: 474JKTimes Cited: 9 Cited Reference Count: 76 Calhorda, MJ Drew, MGB Felix, V Fonseca, LP Gamelas, CA Godinho, SSMC Goncalves, IS Hunstock, E Lopes, JP Parola, AJ Pina, F Romao, CC Santos, AG Felix, Vitor/D-8440-2011; Pina, Fernando/C-8161-2011; Romao, Carlos /D-1162-2010; Parola, A. Jorge/F-4048-2010; Goncalves, Isabel/H-3742-2013 Romao, Carlos /0000-0001-5061-3743; Parola, A. Jorge/0000-0002-1333-9076; Elsevier science sa Lausanne

Protonation and Zn(II) coordination by dipyridine-containing macrocycles with different molecular architecture. A case of pH-controlled metal jumping outside-inside the macrocyclic cavity - Lodeiro, C.; A. J. Parola; Pina, F; Bazzicalupi, C; Bencini, A; Bianchi, A; Giorgi, C; Masotti, A.; Valtancoli, B (Inorganic Chemistry, 2001)
The synthesis of the macrocyclic ligand 4,4 '-(2,5,8, 11,14-pentaaza[15])-2,2 ' -bipyridylophane (L3), which contains a pentaamine chain linking the 4,4 ' -positions of a 2,2 ' -dipyridine moiety, is reported. Protonation and Zn(II) complexation by L3 and by macrocycle L2, containing the same pentaamine chain connecting the 6,6 ' -positions of 2,2 ' -dipyridine, were studied by means of potentiometric, UV-vis, and fluorescent emission measurements. While in L2 all the nitrogen donor atoms are convergent inside the macrocyclic cavity, in L3 the heteroaromatic nitrogen atoms are located outside. Both ligands form mono- and dinuclear Zn(II) complexes in aqueous solution. In the mononuclear Zn(II) complexes with L2, the metal is coordinated inside the macrocyclic cavity, bound to the heteroaromatic nitrogen donors and three amine groups of the aliphatic chain. As shown by the crystal structure of the [ZnL2](2+) complex, the two benzylic nitrogens are not coordinated and facile protonation of the complex takes place at slightly acidic pH values. Considering the mononuclear [ZnL3](2+) complex, the metal is encapsulated inside the cavity, not coordinated by the dipyridine unit. Protonation of the complex occurs on the aliphatic polyamine chain and gives rise to translocation of the metal outside the cavity, bound to the, heteroaromatic nitrogens.
ISI Document Delivery No.: 443EQTimes Cited: 47 Cited Reference Count: 67 Lodeiro, C Parola, AJ Pina, F Bazzicalupi, C Bencini, A Bianchi, A Giorgi, C Masotti, A Valtancoli, B Masotti, Andrea/D-5306-2009; Pina, Fernando/C-8161-2011; Lodeiro, Carlos/B-4793-2013; Parola, A. Jorge/F-4048-2010 Masotti, Andrea/0000-0002-2511-5088; Lodeiro, Carlos/0000-0001-5582-5446; Parola, A. Jorge/0000-0002-1333-9076 Amer chemical soc Washington

Cellulose-based composite films - Borges, João Paulo; Godinho, Maria Helena; Martins, Assis Farinha; Trindade, Ana Catarina; Belgacem, Mohamed Naceur (Mechanics of composite materials, 2001)
The mechanical and optical properties of cellulose-based composite films are investigated.It is shown that the use of toluene diisocyanate as a coupling agent and Avicel fibers as reinforcing elements give films with the highest mechanical characteristics. Using differential scanning calorimetry, it is also found that the glass transition temperature Tg of all the materials studied is below the room temperature and that the Tg increased with cross-linking and introduction of Avicel.

Complementation of the essential peptidoglycan transpeptidase function of penicillin-binding protein (PBP) 2 by the drug resistance protein PBP2A in Staphylococcus aureus - M. G. Pinho; S. R. Filipe; H. de Lencastre; A. Tomasz (J. Bacteriol, 2001)

Conservação Energética em Edifícios de Habitação - Amado, Miguel (6ª Conferência Nacional sobre a Qualidade do Ambiente, 2001)

Conservação Energética em edifícios de habitação e o nível de conforto ambiental - Amado, Miguel (2001)

Effect of Hydrogen-Bond Networks in Controlling Reduction Potentials in Desulfovibrio vulgaris (Hildenborough) Cytochrome c3 Probed by Site-Specific Mutagenesis - Salgueiro, C.A.; da Costa, Patrícia N.; Turner, David L.; Ana C. Messias; van Dongen, Walter M. A. M.; Saraiva, Lígia M.; Xavier, António V. (Biochemistry, 2001)
Cytochromes c3 isolated from Desulfovibrio spp. are periplasmic proteins that play a central role in energy transduction by coupling the transfer of electrons and protons from hydrogenase. Comparison between the oxidized and reduced structures of cytochrome c3 isolated from Desulfovibrio vulgaris (Hildenborough) show that the residue threonine 24, located in the vicinity of heme III, reorients between these two states [Messias, A. C., Kastrau, D. H. W., Costa, H. S., LeGall, J., Turner, D. L., Santos, H., and Xavier, A. V. (1998) J. Mol. Biol. 281, 719−739]. Threonine 24 was replaced with valine by site-directed mutagenesis to elucidate its effect on the redox properties of the protein. The NMR spectra of the mutated protein are very similar to those of the wild type, showing that the general folding and heme core architecture are not affected by the mutation. However, thermodynamic analysis of the mutated cytochrome reveals a large alteration in the microscopic reduction potential of heme III (75 and 106 mV for the protonated forms of the fully reduced and oxidized states, respectively). The redox interactions involving this heme are also modified, while the remaining heme−heme interactions and the redox−Bohr interactions are less strongly affected. Hence, the order of oxidation of the hemes in the mutated cytochrome is different from that in the wild type, and it has a higher overall affinity for electrons. This is consistent with the replacement of threonine 24 by valine preventing the formation of a network of hydrogen bonds, which stabilizes the oxidized state. The mutated protein is unable to perform a concerted two-electron step between the intermediate oxidation stages, 1 and 3, which can occur in the wild-type protein. Thus, replacing a single residue unbalances the global network of cooperativities tuned to control thermodynamically the directionality of the stepwise electron transfer and may affect the functionality of the protein.

Functional analysis of Streptococcus pneumoniae MurM reveals the region responsible for its specificity in the synthesis of peptidoglycan branched peptides - S. R. Filipe; E. Severina; A. Tomasz (J. Biol. Chem, 2001)

Involuntary Rhythmic Leg Movements Time-Locked With the Respiratory Cycle - Alberto Leal; Eulália Calado (Pediatr Neurol, 2001)

New bio-composites based on short fibre reinforced hydroxypropylcellulose films - Borges, João Paulo; Godinho, Maria Helena; Belgacem, Mohamed Naceur; Martins, Assis Farinha (Composite interfaces, 2001)
The present work deals with the preparation and the characterisation of solid films, having thickness between 15 and 40 μm, prepared with hydroxypropyl cellulose (HPC) and different amounts of commercial cellulose fibres (0; 0.5; 10 and 15% w/w) (system A) and with 1,4-butyl diisocyanate (BDI), as a cross-linking agent (system B). Before the preparation of these films, the surface energy of cellulose fibres, as such and after purification with different solvents, was determined by Inverse Gas Chromatography (IGC), which gave the values of dispersive energy and acid-base properties of their surfaces. The tensile and photo-elastic properties of the solid films obtained were studied. There were no significant changes in Young's modulus between the two systems. However, as expected, the values of the elongation and those of the fracture stress were systematically higher for system B in comparison with system A. The same trend was found for the birefringence, measured at the same elongation values for the films obtained from both systems.

NMR structure of the haem core of a novel tetrahaem cytochrome isolated from Shewanella frigidimarina: identification of the haem-specific axial ligands and order of oxidation - Miguel Pessanha; Brennan, Lorraine; Xavier, António V.; Cuthbertson, Pauline M; Graeme A. Reid; Stephen K. Chapman; Turner, David L.; Salgueiro, C.A. (FEBS Letters, 2001)
The tetrahaem cytochrome isolated during anaerobic growth of Shewanella frigidimarina NCIMB400 is a small protein (86 residues) involved in electron transfer to Fe(III), which can be used as a terminal respiratory oxidant by this bacterium. A 3D solution structure model of the reduced form of the cytochrome has been determined using NMR data in order to determine the relative orientation of the haems. The haem core architecture of S. frigidimarina tetrahaem cytochrome differs from that found in all small tetrahaem cytochromes c3 so far isolated from strict anaerobes, but has some similarity to the N-terminal cytochrome domain of flavocytochrome c3 isolated from the same bacterium. NMR signals obtained for the four haems of S. frigidimarina tetrahaem cytochrome at all stages of oxidation were cross-assigned to the solution structure using the complete network of chemical exchange connectivities. Thus, the order in which each haem in the structure becomes oxidised was determined.

Patologias do edifício do Grande Auditório da FCT – CAMPUS DA FACULDADE DE CIÊNCIAS E TECNOLOGIA - Amado, Miguel (Gabinete de Planeamento Físico e Gestão Ambiental da Faculdade de Ciências e Tecnologia da UNL, 2001)

The role of the murMN operon in penicillin resistance and antibiotic tolerance of Streptococcus pneumoniae - S. R. Filipe; E. Severina; A. Tomasz (Microb. Drug Resist, 2001)

Temperature Calibration in dielectric measurements - J.F. Mano; S. Lanceros-Méndez; A. M. Nunes; M. Dionísio (Journal of Thermal Analysis and Calorimetry, 2001)

Time-Frequency and Time-Scale Characterisation of the Beat-by-Beat High-Resolution Electrocardiogram - Batista, A.G.; Rodrigues, J.M.; Ortigueira, M.D. (Sixth Portuguese Conference on Biomedical Engineering Proceedings, 2001)
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Book review of Alice R. P. Abreu (org.): Flexible production and economic governance in Latin America - António Moniz (2001)
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Keywords: actual President of ISA-RC30 “Sociology of Work”; but also from Argentina and Chile. All these papers are structured around the three main parts of the book: the first one is on the productive re-structuring and inter-firm relations in automotive production chain; held in September 1997 also in Rio de Janeiro (Brazil). In this book are published papers presented on Latin American case studies; is a follow up of an international seminar on the same theme of the title; mostly from Brazil and Mexico; the second one on productive re-structuring and flexibility and the third one on entrepreneurs and entrepreneurial associations.; This book organized by Alice Abreu

A cooperação entre equipas de trabalho em empresas em rede: vantagens para o desenvolvimento regional [Workteam Co-operation in Networked Companies: regional development advantages] - António Moniz (University Library of Munich, Germany, 2001)
Working teams in enterprise environment are considered as the most advanced forms of work organisation. This means the forms that can improve productivity quality of working life. Nevertheless, it prevails a slow development and dissemination of these advanced organisational forms in European companies. The reason for that lays in a complex linkage factors from social values to the economical pressures. But other factors are also related to the national systems of education training, to the different systems of industrial relations and technology policy.
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Keywords: enterprise networks; Europe; innovation; labour; productivity; work organization

Coordination properties of a polyamine cryptand with two different binding moieties. A case of a pH-modulated antenna device based on a new Eu(III) cryptate complex - Bazzicalupi, C; Bencini, A; Berni, E; Bianchi, A; Giorgi, C; Fusi, V.; Valtancoli, B; Lodeiro, C.; Roque, A.; Pina, F (Inorganic Chemistry, 2001)
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Times Cited: 16 Pina, Fernando/C-8161-2011; Fusi, Vieri/H-1006-2012; Lodeiro, Carlos/B-4793-2013; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, C4O/M-5599-2013; REQUIMTE, UCIBIO/N-9846-2013; REQUIMTE, LAQV/N-9835-2013 Pina, Fernando/0000-0001-8529-6848; Lodeiro, Carlos/0000-0001-5582-5446; 16

Dissimilatory Nitrate Reductase - Romão, M.J.; Dias, J. M.; Moura, I. (Handbook of Metalloproteins, 2001)
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Electrochemical studies of rubredoxin from Desulfovibrio vulgaris at modified electrodes - Correia dos Santos, M. M.; Paes de Sousa, P. M.; Simões Gonçalves, M. L.; Ascenso, C.; Moura, I.; Moura, J. J. G. (Journal of Electroanalytical Chemistry, 2001)
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Keywords: Modified macro- and micro-electrodes; Rubredoxin; Unmediated electrochemistry

Information Society, Work and the Generation of New Forms of Social Exclusion (SOWING): National Report (Portugal) - António Moniz; Gomes, Cláudia; Machado, Tiago; Urze, Paula (University Library of Munich, Germany, 2001)
The choice over the Portuguese case studies was based on the sample constructed for the application of the firm questionnaires, during the second year of the SOWING project, 1999. This sample was fulfilled of firms among several activity sectors: textile, manufacturing, electronics, transports and software industry, based on NACE – codes (2 – digit level). Thus, we agreed to include in a new database the remaining questionnaires and construct a sample with 113 observations. Concerning the organisational change we make a distinction of three categories of change. First we analyse changes taking place at the inter-firm level (outsourcing, subcontracting, geographic relocation), followed by changes at the organisational level (deconcentration/decentralisation, reduction of hierarchical levels, introduction of cost and profit centres). The third kind of changes analysed will be those taking place at the workplace level (job enlargement/enrichment, changing character of work, work load). The Portuguese studied companies presents a relative uniform pattern considering the variables social competencies, practical knowledge, responsibility and specialized professional qualifications.
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Keywords: industry; information technologies; organisation; qualification; Work

Internal dynamics of poly(methylphenylsiloxane) chains as revealed by picosecond time resolved fluorescence - Dias, FB; J. C. Lima; Pierola, IF; Horta, A; Macanita, AL (Journal of Physical Chemistry a, 2001)
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Ionic liquids as recyclable reaction media for the tetrahydropyranylation of alcohols - Branco, L. C.; C. A. M. Afonso (Tetrahedron, 2001)
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Times Cited: 67 Branco, Luis/C-6426-2013; REQUIMTE, PHOTO/M-4229-2013; Mateus Afonso, Carlos Alberto/M-7833-2013; REQUIMTE, LAQV/N-9835-2013; iMed.ULisboa, iMed.ULisboa/C-6292-2014; iMed.ULisboa, BioOrgChem /B-4209-2014 Branco, Luis/0000-0003-2520-1151; Mateus Afonso, Carlos Alberto/0000-0002-7284-5948; 69

Micelle effect on ground and excited state proton transfer reactions involving the 4-methyl-7-hydroxyflavylium cation - Pina, F; Melo, MJ; Alves, S; Ballardini, R; Maestri, M; Passaniti, P. (New Journal of Chemistry, 2001)
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Times Cited: 19 Melo, Maria Joao/C-8594-2011; Pina, Fernando/C-8161-2011; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Melo, Maria Joao/0000-0001-7393-6801; Pina, Fernando/0000-0001-8529-6848; 19

Mossbauer characterization of the iron-sulfur clusters in Desulfovibrio vulgaris hydrogenase - Pereira, A. S.; Tavares, P; Moura, I.; Moura, J. J. G.; Huynh, B. H. (Journal Of The American Chemical Society, 2001)
The periplasmic hydrogenase of Desulfovibrio vulgaris (Hildenbourough) is an all Fe-containing hydrogenase. It contains two ferredoxin type [4Fe-4S] clusters, termed the F clusters, and a catalytic H cluster. Recent X-ray crystallographic studies on two Fe hydrogenases revealed that the H cluster is composed of two sub-clusters, a [4Fe-4S] cluster ([4Fe-4S]H) and-a binuclear Fe cluster ([2Fe]H), bridged by a cysteine sulfur. The aerobically purified D. vulgaris hydrogenase is stable in air. It is inactive and requires reductive activation. Upon reduction, the enzyme becomes sensitive to O(2) indicating that the reductive activation process is irreversible. Previous EPR investigations showed that upon reoxidation (under argon) the H cluster exhibits a rhombic EPR signal that is not seen in the as-purified enzyme, suggesting a conformational change in association with the reductive activation. For the purpose of gaining more information on the electronic properties of this unique H cluster and to understand further the reductive activation process, variable-temperature and variable-field Mossbauer spectroscopy has been used to characterize the Fe-S clusters in D. vulgaris hydrogenase poised at different redox states generated during a reductive titration, and in the GO-reacted enzyme. The data were successfully decomposed into spectral components corresponding to the F and H clusters,and characteristic parameters describing the electronic and magnetic properties of the F and H clusters were obtained. Consistent with the X-ray crystallographic results, the spectra of the H cluster can be understood as originating from an exchange coupled [4Fe-4S] - [2Fe] system. In particular, detailed analysis of the data reveals that the reductive activation begins with reduction of the [4Fe-4S]H cluster from the 2+ to the If state, followed by transfer of the reducing equivalent from the [4Fe-4S]H subcluster to the binuclear [2Fe]H subcluster. The results also reveal that binding of exogenous CO to the H cluster affects significantly the exchange coupling between the [4Fe-4S]H and the [2Fe]H subclusters. Implication of such a CO binding effect is discussed.
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Open-chain polyamine ligands bearing an anthracene unit - Chemosensors for logic operations at the molecular level - Alves, S; Pina, F; Albelda, MT; Garcia-Espana, E; Soriano, C; Luis, SV (European Journal of Inorganic Chemistry, 2001)
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Times Cited: 69 Pina, Fernando/C-8161-2011; Albelda, M. Teresa/J-5929-2012; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Pina, Fernando/0000-0001-8529-6848; 69

Palladium(II)-promoted aziridination of olefins with bromamine T as the nitrogen transfer reagent - A.M.M. Antunes; Marto, SJL; Branco, P. S.; Prabhakar, S.; A. M. Lobo (CHEMICAL COMMUNICATIONS, 2001)
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Photochemical- and pH-switching properties of a new photoelastic ligand based upon azobenzene. Basicity and anion binding - Bencini, A; Bianchi, A; Giorgi, C; Romagnoli, E.; Lodeiro, C.; Saint-Maurice, A.; Pina, F; Valtancoli, B (Supramolecular Chemistry, 2001)
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Times Cited: 7 Pina, Fernando/C-8161-2011; Lodeiro, Carlos/B-4793-2013; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, C4O/M-5599-2013; REQUIMTE, UCIBIO/N-9846-2013; REQUIMTE, LAQV/N-9835-2013 Pina, Fernando/0000-0001-8529-6848; Lodeiro, Carlos/0000-0001-5582-5446; 7

Photoinduced electron transfer in non-aqueous microemulsions - Costa, S. M. B.; Lopez-Cornejo, P.; Togashi, D. M.; Laia, C. A. T. (Journal of Photochemistry and Photobiology a-Chemistry, 2001)
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Times Cited: 16 Laia, Cesar/C-8651-2011; Costa, Silvia/H-7766-2012; REQUIMTE, PHOTO/M-4229-2013; Costa, Silvia/M-8085-2013; REQUIMTE, LAQV/N-9835-2013 Laia, Cesar/0000-0001-6410-6072; Costa, Silvia/0000-0002-6978-8903; 16

Polyamine linear chains bearing two identical terminal aromatic units. Evidence for a photo induced bending movement - Bernardo, M. A.; Alves, S; Pina, F; de Melo, JS; Albelda, MT; Garcia-Espana, E; Llinares, J. M.; Soriano, C; Luis, SV (Supramolecular Chemistry, 2001)
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Times Cited: 21 Pina, Fernando/C-8161-2011; Seixas de Melo, Sergio/A-5388-2008; Albelda, M. Teresa/J-5929-2012; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Pina, Fernando/0000-0001-8529-6848; Seixas de Melo, Sergio/0000-0001-9708-5079; 21

Polyamines containing naphthyl groups as pH-regulated molecular machines driven by light - Albelda, MT; Bernardo, M. A.; Diaz, P; Garcia-Espana, E; de Melo, JS; Pina, F; Soriano, C; Santiago, V. L. E. (Chemical Communications, 2001)
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Times Cited: 46 Pina, Fernando/C-8161-2011; Seixas de Melo, Sergio/A-5388-2008; Albelda, M. Teresa/J-5929-2012; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Pina, Fernando/0000-0001-8529-6848; Seixas de Melo, Sergio/0000-0001-9708-5079; 46

Proteómica: a Interface entre a Biologia Molecular e a Biochemistry de Proteínas - Almeida, G.; C. Rodrigues; Lampreia, J. (Bol. Soc. Port. Química, 2001)
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Purification, crystallization and identification by X-ray analysis of a prostate kallikrein from horse seminal plasma - Carvalho, A. L.; Dias, J. M.; Sanz, L.; Romero, A.; Calvete, J. J.; Romao, M. J. (Acta Crystallographica Section D-Biological Crystallography, 2001)
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Times Cited: 2 Romao, Maria/A-4115-2013; Carvalho, Ana Luisa/G-5638-2011; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Carvalho, Ana Luisa/0000-0002-3824-0240; 8 2

Rational reprogramming of the R2 subunit of Escherichia coli ribonucleotide reductase into a self-hydroxylating monooxygenase - Baldwin, J; Voegtli, WC; Khidekel, N; Moenne-Loccoz, P; Krebs, C.; Pereira, A. S.; Ley, BA; Huynh, B. H.; Loehr, T. M.; Riggs-Gelasco, PJ; Rosenzweig, AC; Bollinger, JM (Journal Of The American Chemical Society, 2001)
The outcome of O-2 activation at the diiron(II) cluster in the R2 subunit of Escherichia coli (class I) ribonucleotide reductase has been rationally altered from the normal tyrosyl radical (Y122)(1) production to self-hydroxylation of a phenylalanine side-chain by two amino acid substitutions that leave intact the (histidine)(2)-(carboxylate)(4) ligand set characteristic of the diiron-carboxylate family. Iron ligand Asp (D) 84 was replaced with Glu (E), the amino acid found in the cognate position of the structurally similar diiron-carboxylate protein, methane monooxygenase hydroxylase (MMOH). We previously showed that this substitution allows accumulation of a mu -1,2-peroxodiiron(III) intermediate,(2 3) which does not accumulate in the wild-type (wt) protein and is probably a structural homologue of intermediate P (H-peroxo) in O-2 activation by MMOH.(4) In addition, the near-surface residue Trp (W) 48 was replaced with Phe (F), blocking transfer of the ``extra'' electron that occurs in wt R2 during formation of the formally Fe(LII)Fe(IV) cluster X.(5-7) Decay of the mu1,2-peroxodiiron(III) complex in R2-W38F/D84E gives an initial brown product, which contains very little YI22(.) and which converts very slowly (t(1/2) similar to 7 h) upon incubation at 0 degreesC to an intensely purple final product. X-ray crystallographic analysis of the purple product indicates that F208 has undergone epsilon -hydroxylation and the resulting phenol has shifted significantly to become st ligand to Fe2 of the diiron cluster. Resonance Raman (RR) spectra of the purple product generated with O-16(2) or O-18(2) show appropriate isotopic sensitivity in bands assigned to O-phenyl and Fe-O-phenyl vibrational modes, confirming that the oxygen of the Fe(III)-phenolate species is derived from Or. Chemical analysis, experiments involving interception of the hydroxylating intermediate with exogenous reductant, and Mossbauer and EXAFS characterization of the brown and purple species establish that F208 hydroxylation occurs during decay of the peroxo complex and formation of the initial brown product. The slow transition to the purple Fe(LII)-phenolate species is ascribed to a ligand rearrangement in which mu -O2- is lost and the F208-derived phenolate coordinates. The reprogramming to F208 monooxygenase requires both amino acid substitutions, as very little epsilon -hydroxyphenylalanine is formed and pathways leading to Y122(.) formation predominate in both R2-D84E and R2-W48F(2-7).
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Solvatochromism and thermochromism of the electronic spectra of an indocarbocyanine dye - Laia, C. A. T.; Costa, S. M. B. (Journal of Molecular Structure, 2001)
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Times Cited: 2 Laia, Cesar/C-8651-2011; Costa, Silvia/H-7766-2012; REQUIMTE, PHOTO/M-4229-2013; Costa, Silvia/M-8085-2013; REQUIMTE, LAQV/N-9835-2013 Laia, Cesar/0000-0001-6410-6072; Costa, Silvia/0000-0002-6978-8903; Si XXVth European Congress on Molecular Spectroscopy Aug 27-sep 01, 2000 Coimbra, portugal 2

Structural studies on Desulfovibrio desulfuricans ATCC 27774 multiheme nitrite reductase - characterization of the subunits - Goncalves, L. M. L.; Cunha, C.; Almeida, G.; Macieira, S.; Costa, C.; Lampreia, J.; Romao, M. J.; Moura, J. J. G.; Moura, I. (Journal of Inorganic Biochemistry, 2001)
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Times Cited: 0 Costa, Cristina/A-9966-2012; Romao, Maria/A-4115-2013; Lampreia, Jorge/A-9927-2012; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013; Almeida, Maria Gabriela/D-8265-2013 Costa, Cristina/0000-0002-8611-9023; Romao, Maria/0000-0002-3004-0543; Lampreia, Jorge/0000-0001-8846-0041; Moura, Isabel/0000-0003-0971-4977; Almeida, Maria Gabriela/0000-0002-4508-7379 0

Structure refinement of the aldehyde oxidoreductase from Desulfovibrio gigas (MOP) at 1.28 angstrom - Rebelo, J. M.; Dias, J. M.; Huber, R.; Moura, J. J. G.; Romao, M. J. (Journal of Biological Inorganic Chemistry, 2001)
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Times Cited: 52 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 54

Substitution of murine ferrochelatase glutamate-287 with glutamine or alanine leads to porphyrin substrate-bound variants - Franco, R.; Pereira, A. S.; Tavares, P; Mangravita, A.; Barber, M. J.; Moura, I.; Ferreira, G. C. (BIOCHEMICAL JOURNAL, 2001)
Ferrochelatase (EC 4.99.1.1) is the terminal enzyme of the haem biosynthetic pathway and catalyses iron chelation into the protoporphyrin IX ring. Glutamate-287 (E287) of murine mature ferrochelatase is a conserved residue in all known sequences of ferrochelatase, is present at the active site of the enzyme, as inferred from the Bacillus subtilis ferrochelatase three-dimensional structure, and is critical for enzyme activity. Substitution of E287 with either glutamine (Q) or alanine (A) yielded variants with lower enzymic activity than that of the wild-type ferrochelatase and with different absorption spectra from the wild-type enzyme. In contrast to the wild-type enzyme, the absorption spectra of the variants indicate that these enzymes, as purified, contain protoporphyrin IX. Identification and quantification of the porphyrin bound to the E287-directed variants indicate that approx. 80% of the total porphyrin corresponds to protoporphyrin IX. Significantly, rapid stopped-flow experiments of the E287A and E287Q Variants demonstrate that reaction with Zn2+ results in the formation of bound Zn-protoporphyrin IX, indicating that the endogenously bound protoporphyrin IX can be used as a substrate. Taken together, these findings suggest that the structural strain imposed by ferrochelatase on the porphyrin substrate as a critical step in the enzyme catalytic mechanism is also accomplished by the E287A and E287Q variants, but without the release of the product. Thus E287 in murine ferrochelatase appears to be critical For the catalytic process by controlling the release of the product.
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Time-Frequency and Time-Scale Characterisation of the Beat-by-Beat High-Resolution Electrocardiogram - Batista, A.G.; Rodrigues, J.M.; Ortigueira, M.D. (Sixth Portuguese Conference on Biomedical Engineering Proceedings, 2001)
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Tungsten-containing formats dehydrogenase from Desulfovibrio gigas: metal identification and preliminary structural data by multi-wavelength crystallography - Raaijmakers, H.; Teixeira, S.; Dias, J. M.; Almendra, M. J.; Brondino, C. D.; Moura, I.; Moura, J. J. G.; Romao, M. J. (Journal of Biological Inorganic Chemistry, 2001)
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Times Cited: 25 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 29

{A cooperação entre equipas de trabalho em empresas em rede: vantagens para o desenvolvimento regional [Workteam Co-operation in Networked Companies: regional development advantages]} - António Moniz (University Library of Munich, Germany, 2001)
Working teams in enterprise environment are considered as the most advanced forms of work organisation. This means the forms that can improve productivity quality of working life. Nevertheless, it prevails a slow development and dissemination of these advanced organisational forms in European companies. The reason for that lays in a complex linkage factors from social values to the economical pressures. But other factors are also related to the national systems of education training, to the different systems of industrial relations and technology policy.
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Keywords: enterprise networks; Europe; innovation; labour; productivity; work organization

{Book review of Alice R. P. Abreu (org.): Flexible production and economic governance in Latin America} - António Moniz (2001)
5937
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Keywords: actual President of ISA-RC30 “Sociology of Work”; but also from Argentina and Chile. All these papers are structured around the three main parts of the book: the first one is on the productive re-structuring and inter-firm relations in automotive production chain; held in September 1997 also in Rio de Janeiro (Brazil). In this book are published papers presented on Latin American case studies; is a follow up of an international seminar on the same theme of the title; mostly from Brazil and Mexico; the second one on productive re-structuring and flexibility and the third one on entrepreneurs and entrepreneurial associations.; This book organized by Alice Abreu

{Information Society, Work and the Generation of New Forms of Social Exclusion (SOWING): National Report (Portugal)} - António Moniz; Gomes, Cláudia; Machado, Tiago; Urze, Paula (University Library of Munich, Germany, 2001)
The choice over the Portuguese case studies was based on the sample constructed for the application of the firm questionnaires, during the second year of the SOWING project, 1999. This sample was fulfilled of firms among several activity sectors: textile, manufacturing, electronics, transports and software industry, based on NACE – codes (2 – digit level). Thus, we agreed to include in a new database the remaining questionnaires and construct a sample with 113 observations. Concerning the organisational change we make a distinction of three categories of change. First we analyse changes taking place at the inter-firm level (outsourcing, subcontracting, geographic relocation), followed by changes at the organisational level (deconcentration/decentralisation, reduction of hierarchical levels, introduction of cost and profit centres). The third kind of changes analysed will be those taking place at the workplace level (job enlargement/enrichment, changing character of work, work load). The Portuguese studied companies presents a relative uniform pattern considering the variables social competencies, practical knowledge, responsibility and specialized professional qualifications.
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Keywords: industry; information technologies; organisation; qualification; Work

{Technological practices in the European auto industry: Exploring cases from Belgium, Germany and Portugal} - António Moniz; Krings, Bettina; Van Hootegem, Geert; Huys, Rik (University Library of Munich, Germany, 2001)
The relation between work organisation and technological practices in auto industry is analysed in this article. The concept of “technological practice” in this sector is used to describe the specific ways of embedding information and communication technology applications into the organizational forms and cultural patterns. This concept was developed with the Sowing project (TSER, DG XII) and that approach included either the shop floor co-operation up to the regionally based networks of companies and supporting institutions. The authors studied different sectors in the automotive firms of different European countries (Germany, Belgium and Portugal): shopfloor and production lines, design and management and the local inter-relationships. It was underlined some evidencies of the different alternatives in terms of technological practices for the same sector. Much of the litterature try to disseminate an idea of a single (and optimum) organisational model for the same type of product. And here, even with the same type of technology, and of product (medium-high range), one can find different models, different cultures, different ways of organising the industrial structure (firms, regional institutions, R&D centres) in the same sector (auto industry).
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Keywords: automobile sector; Information and Communication Technologies; technological practice; work organisa

The qualitative probing of hydrogen bond strength by diffusion-ordered NMR spectroscopy - Kapur, GS; E. J. Cabrita; Berger, S (Tetrahedron Letters, 2000)
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Redox potential measurements of the Mycobacterium tuberculosis heme protein KatG and the isoniazid-resistant enzyme KatG(S315T): insights into isoniazid activation - Wengenack, N. L.; Lopes, H.; Kennedy, M. J.; Tavares, P; Pereira, A. S.; Moura, I.; Moura, J. J.; Rusnak, F. (Biochemistry, 2000)
Mycobacterium tuberculosis KatG is a multifunctional heme enzyme responsible for activation of the antibiotic isoniazid. A KatG(S315T) point mutation is found in >50% of isoniazid-resistant clinical isolates. Since isoniazid activation is thought to involve an oxidation reaction, the redox potential of KatG was determined using cyclic voltammetry, square wave voltammetry, and spectroelectrochemical titrations. Isoniazid activation may proceed via a cytochrome P450-like mechanism. Therefore, the possibility that substrate binding by KatG leads to an increase in the heme redox potential and the possibility that KatG(S315T) confers isoniazid resistance by altering the redox potential were examined. Effects of the heme spin state on the reduction potentials of KatG and KatG(S315T) were also determined. Assessment of the Fe(3+)/Fe(2+) couple gave a midpoint potential of ca. -50 mV for both KatG and KatG(S315T). In contrast to cytochrome P450s, addition of substrate had no significant effect on either the KatG or KatG(S315T) redox potential. Conversion of the heme to a low-spin configuration resulted in a -150 to -200 mV shift of the KatG and KatG(S315T) redox potentials. These results suggest that isoniazid resistance conferred by KatG(S315T) is not mediated through changes in the heme redox potential. The redox potentials of isoniazid were also determined using cyclic and square wave voltammetry, and the results provide evidence that the ferric KatG and KatG(S315T) midpoint potentials are too low to promote isoniazid oxidation without formation of a high-valent enzyme intermediate such as compounds I and II or oxyferrous KatG.
0006-2960 (Print)0006-2960 (Linking)Journal Article
Keywords: *Bacterial Proteins; Amino Acid Substitution/genetics; Antitubercular Agents/*pharmacology; Drug Resistance, Microbial; Enzyme Activation/drug effects/genetics; Free Radicals/metabolism; Hemeproteins/genetics/*metabolism; Isoniazid/*pharmacology; Mycobacterium tuberculosis/drug effects/*enzymology; Oxidation-Reduction/drug effects; Peroxidases/genetics/*metabolism; Potentiometry; Serine/genetics; Sodium Cyanide/pharmacology; Spectrophotometry; Threonine/genetics

Neelaredoxin, an iron-binding protein from the syphilis spirochete, Treponema pallidum, is a superoxide reductase - Jovanovic, T.; Ascenso, C.; Hazlett, K. R.; Sikkink, R.; Krebs, C.; Litwiller, R.; Benson, L. M.; Moura, I.; Moura, J. J.; Radolf, J. D.; Huynh, B. H.; Naylor, S.; et al. (J Biol Chem, 2000)
Treponema pallidum, the causative agent of venereal syphilis, is a microaerophilic obligate pathogen of humans. As it disseminates hematogenously and invades a wide range of tissues, T. pallidum presumably must tolerate substantial oxidative stress. Analysis of the T. pallidum genome indicates that the syphilis spirochete lacks most of the iron-binding proteins present in many other bacterial pathogens, including the oxidative defense enzymes superoxide dismutase, catalase, and peroxidase, but does possess an orthologue (TP0823) for neelaredoxin, an enzyme of hyperthermophilic and sulfate-reducing anaerobes shown to possess superoxide reductase activity. To analyze the potential role of neelaredoxin in treponemal oxidative defense, we examined the biochemical, spectroscopic, and antioxidant properties of recombinant T. pallidum neelaredoxin. Neelaredoxin was shown to be expressed in T. pallidum by reverse transcriptase-polymerase chain reaction and Western blot analysis. Recombinant neelaredoxin is a 26-kDa alpha(2) homodimer containing, on average, 0.7 iron atoms/subunit. Mossbauer and EPR analysis of the purified protein indicates that the iron atom exists as a mononuclear center in a mixture of high spin ferrous and ferric oxidation states. The fully oxidized form, obtained by the addition of K(3)(Fe(CN)(6)), exhibits an optical spectrum with absorbances at 280, 320, and 656 nm; the last feature is responsible for the protein's blue color, which disappears upon ascorbate reduction. The fully oxidized protein has a A(280)/A(656) ratio of 10.3. Enzymatic studies revealed that T. pallidum neelaredoxin is able to catalyze a redox equilibrium between superoxide and hydrogen peroxide, a result consistent with it being a superoxide reductase. This finding, the first description of a T. pallidum iron-binding protein, indicates that the syphilis spirochete copes with oxidative stress via a primitive mechanism, which, thus far, has not been described in pathogenic bacteria.
0021-9258 (Print)0021-9258 (Linking)Journal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.
Keywords: Amino Acid Sequence; Bacterial Proteins/*metabolism; Carrier Proteins/*metabolism; Cloning, Molecular; Electron Spin Resonance Spectroscopy; Iron-Binding Proteins; Molecular Sequence Data; Oxidoreductases/*metabolism; Recombinant Proteins/isolation & purification; Superoxide Dismutase/metabolism; Transferrin-Binding Proteins; Treponema pallidum/*metabolism

I&D, inovação e fomento de emprego: ideias chave de uma relação [R&D, innovation and job creation: key-ideas of a relation] - António Moniz; Casaca, Sara; Bairrada, Mário; Moreno, Conceição (University Library of Munich, Germany, 2000)
The increasing degree of inter-dependency and integration of economy at a global scale motivated the globalization of scientific and technological knowledge from manufacturing, to the marketing and the strategic management of companies that act as protagonists of these processes. Globalization of markets is inter-connected with intensive incorporation of knowledge in economy. In this sense the continuous evolution of the manufacturing industry structure for an increased high intensity technology elements, namely information and communication technologies, implies that these industrial sub-sectors are conditioning the global performance of economy, the productivity gains and as a consequence the levels of economical and employment growth. This study on “R&D, Innovation and Employment Creation” for the Portuguese Observatory of Employment and Vocational Training (IEFP-MQE) is updating information on the articulation between Job creation, innovation, technology, and R&D. A diagnosis of the state of national S&T research and of processes of technological and organizational innovation was made, as well of the interfacing structures between the knowledge sector and the economical activities. Were made 12 case studies at companies with some innovation activities, interviews to unionists and to Professional associations, and public offices. Was made also a survey to a selected sample of technological infrastructures.
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Keywords: innovation; job creation; knowledge; research & development; strategic management; technological inf

{I&D, inovação e fomento de emprego: ideias chave de uma relação [R&D, innovation and job creation: key-ideas of a relation]} - António Moniz; Casaca, Sara; Bairrada, Mário; Moreno, Conceição (University Library of Munich, Germany, 2000)
The increasing degree of inter-dependency and integration of economy at a global scale motivated the globalization of scientific and technological knowledge from manufacturing, to the marketing and the strategic management of companies that act as protagonists of these processes. Globalization of markets is inter-connected with intensive incorporation of knowledge in economy. In this sense the continuous evolution of the manufacturing industry structure for an increased high intensity technology elements, namely information and communication technologies, implies that these industrial sub-sectors are conditioning the global performance of economy, the productivity gains and as a consequence the levels of economical and employment growth. This study on “R&D, Innovation and Employment Creation” for the Portuguese Observatory of Employment and Vocational Training (IEFP-MQE) is updating information on the articulation between Job creation, innovation, technology, and R&D. A diagnosis of the state of national S&T research and of processes of technological and organizational innovation was made, as well of the interfacing structures between the knowledge sector and the economical activities. Were made 12 case studies at companies with some innovation activities, interviews to unionists and to Professional associations, and public offices. Was made also a survey to a selected sample of technological infrastructures.
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Keywords: innovation; job creation; knowledge; research & development; strategic management; technological inf

Work organisation in industry: Practices of use of IT in Portugal - António Moniz (University Library of Munich, Germany, 2000)
The innovation in the employment behaviour is particularly clear in what concerns the sector composition of the employment changes: the persistent decrease of the industrial employment (even if within a framework of some recovery of the industrial product), a slight expansion in the employment on the tertiary sector, and an important growth of the employment in the construction and public works activities. We can identify then a considerable sector mutation concerning the industrial and the services sectors but also a growth of the atypical forms of employment. Portugal continues not being able to respond to the labour market needs (improvement in the education sector is critical to the catch-up process). The Portuguese cases studied point out to organisational changes supported by ICT, but not determined/induced by it. For most of the changes that were recently developed, ICT had an important role. We didn’t found explicit technological determinism in the relation between ICT and social exclusion.
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Keywords: employment; Information Society; technology determinism

{Work organisation in industry: Practices of use of IT in Portugal} - António Moniz (University Library of Munich, Germany, 2000)
The innovation in the employment behaviour is particularly clear in what concerns the sector composition of the employment changes: the persistent decrease of the industrial employment (even if within a framework of some recovery of the industrial product), a slight expansion in the employment on the tertiary sector, and an important growth of the employment in the construction and public works activities. We can identify then a considerable sector mutation concerning the industrial and the services sectors but also a growth of the atypical forms of employment. Portugal continues not being able to respond to the labour market needs (improvement in the education sector is critical to the catch-up process). The Portuguese cases studied point out to organisational changes supported by ICT, but not determined/induced by it. For most of the changes that were recently developed, ICT had an important role. We didn’t found explicit technological determinism in the relation between ICT and social exclusion.
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Keywords: employment; Information Society; technology determinism

Gene sequence and crystal structure of the aldehyde oxidoreductase from Desulfovibrio desulfuricans ATCC 27774 - Rebelo, J.; Macieira, S.; Dias, J. M.; Huber, R.; Ascenso, C. S.; Rusnak, F.; Moura, J. J.; Moura, I.; Romao, M. J. (J Mol Biol, 2000)
The aldehyde oxidoreductase (MOD) isolated from the sulfate reducer Desulfovibrio desulfuricans (ATCC 27774) is a member of the xanthine oxidase family of molybdenum-containing enzymes. It has substrate specificity similar to that of the homologous enzyme from Desulfovibrio gigas (MOP) and the primary sequences from both enzymes show 68 % identity. The enzyme was crystallized in space group P6(1)22, with unit cell dimensions of a=b=156.4 A and c=177.1 A, and diffraction data were obtained to beyond 2.8 A. The crystal structure was solved by Patterson search techniques using the coordinates of the D. gigas enzyme. The overall fold of the D. desulfuricans enzyme is very similar to MOP and the few differences are mapped to exposed regions of the molecule. This is reflected in the electrostatic potential surfaces of both homologous enzymes, one exception being the surface potential in a region identifiable as the putative docking site of the physiological electron acceptor. Other essential features of the MOP structure, such as residues of the active-site cavity, are basically conserved in MOD. Two mutations are located in the pocket bearing a chain of catalytically relevant water molecules. As deduced from this work, both these enzymes are very closely related in terms of their sequences as well as 3D structures. The comparison allowed confirmation and establishment of features that are essential for their function; namely, conserved residues in the active-site, catalytically relevant water molecules and recognition of the physiological electron acceptor docking site.
0022-2836 (Print)0022-2836 (Linking)Comparative StudyJournal ArticleResearch Support, Non-U.S. Gov't
Keywords: Aldehyde Oxidoreductases/*chemistry/*genetics/metabolism; Amino Acid Sequence; Binding Sites; Conserved Sequence; Crystallization; Crystallography, X-Ray; Cytosine Nucleotides/metabolism; Desulfovibrio/*enzymology/genetics; Dimerization; Hydrogen Bonding; Iron-Sulfur Proteins/chemistry/genetics/metabolism; Models, Molecular; Molecular Sequence Data; Mutation/genetics; Oxidation-Reduction; Protein Structure, Secondary; Pterins/metabolism; Sequence Alignment; Static Electricity; Structure-Activity Relationship; Water/metabolism

Analysis of the electron paramagnetic resonance properties of the [2Fe-2S]1+ centers in molybdenum enzymes of the xanthine oxidase family: assignment of signals I and II - Caldeira, J.; Belle, V.; Asso, M.; Guigliarelli, B.; Moura, I.; Moura, J. J.; Bertrand, P. (Biochemistry, 2000)
Molybdoenzymes of the xanthine oxidase family contain two [2Fe-2S](1+,2+) clusters that are bound to the protein by very different cysteine motifs. In the X-ray crystal structure of Desulfovibrio gigas aldehyde oxidoreductase, the cluster ligated by a ferredoxin-type motif is close to the protein surface, whereas that ligated by an unusual cysteine motif is in contact with the molybdopterin [Romao, M. J., Archer, M., Moura, I., Moura, J. J. G., LeGall, J., Engh, R., Schneider, M., Hof, P., and Huber, R. (1995) Science 270, 1170-1176]. These two clusters display distinct electron paramagnetic resonance (EPR) signals: the less anisotropic one, called signal I, is generally similar to the g(av) approximately 1.96-type signals given by ferredoxins, whereas signal II often exhibits anomalous properties such as very large g values, broad lines, and very fast relaxation properties. A detailed comparison of the temperature dependence of the spin-lattice relaxation time and of the intensity of these signals in D. gigas aldehyde oxidoreductase and in milk xanthine oxidase strongly suggests that the peculiar EPR properties of signal II arise from the presence of low-lying excited levels reflecting significant double exchange interactions. The issue raised by the assignment of signals I and II to the two [2Fe-2S](1+) clusters was solved by using the EPR signal of the Mo(V) center as a probe. The temperature dependence of this signal could be quantitatively reproduced by assuming that the Mo(V) center is coupled to the cluster giving signal I in xanthine oxidase as well as in D. gigas aldehyde oxidoreductase. This demonstrates unambiguously that, in both enzymes, signal I arises from the center which is closest to the molybdenum cofactor.
0006-2960 (Print)0006-2960 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Aldehyde Oxidoreductases/chemistry; Animals; Cattle; Desulfovibrio/enzymology; Electron Spin Resonance Spectroscopy; Iron-Sulfur Proteins/*chemistry; Liver/enzymology; Metalloproteins/chemistry; Milk/enzymology; Molybdenum/*chemistry; Rabbits; Spin Labels; Temperature; Turkeys; Xanthine Oxidase/*chemistry

Heteronuclear NMR and soft docking: an experimental approach for a structural model of the cytochrome c553-ferredoxin complex - Morelli, X.; Dolla, A.; Czjzek, M.; Palma, P. N.; Blasco, F.; Krippahl, L.; Moura, J. J.; Guerlesquin, F. (Biochemistry, 2000)
The combination of docking algorithms with NMR data has been developed extensively for the studies of protein-ligand interactions. However, to extend this development for the studies of protein-protein interactions, the intermolecular NOE constraints, which are needed, are more difficult to access. In the present work, we describe a new approach that combines an ab initio docking calculation and the mapping of an interaction site using chemical shift variation analysis. The cytochrome c553-ferredoxin complex is used as a model of numerous electron-transfer complexes. The 15N-labeling of both molecules has been obtained, and the mapping of the interacting site on each partner, respectively, has been done using HSQC experiments. 1H and 15N chemical shift analysis defines the area of both molecules involved in the recognition interface. Models of the complex were generated by an ab initio docking software, the BiGGER program (bimolecular complex generation with global evaluation and ranking). This program generates a population of protein-protein docked geometries ranked by a scoring function, combining relevant stabilization parameters such as geometric complementarity surfaces, electrostatic interactions, desolvation energy, and pairwise affinities of amino acid side chains. We have implemented a new module that includes experimental input (here, NMR mapping of the interacting site) as a filter to select the accurate models. Final structures were energy minimized using the X-PLOR software and then analyzed. The best solution has an interface area (1037.4 A2) falling close to the range of generally observed recognition interfaces, with a distance of 10.0 A between the redox centers.
0006-2960 (Print)0006-2960 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: *Models, Molecular; Amino Acid Sequence; Base Sequence; Computer Simulation; Cytochrome c Group/*chemistry/genetics/metabolism; Desulfovibrio vulgaris/enzymology; Electron Transport; Ferredoxins/*chemistry/genetics/metabolism; Molecular Sequence Data; Nuclear Magnetic Resonance, Biomolecular/methods; Peptide Mapping; Sulfur-Reducing Bacteria/chemistry

A novel type of catalytic copper cluster in nitrous oxide reductase - Brown, K.; Tegoni, M.; Prudencio, M.; Pereira, A. S.; Besson, S.; Moura, J. J.; Moura, I.; Cambillau, C. (Nat Struct Biol, 2000)
Nitrous oxide (N20) is a greenhouse gas, the third most significant contributor to global warming. As a key process for N20 elimination from the biosphere, N20 reductases catalyze the two-electron reduction of N20 to N2. These 2 x 65 kDa copper enzymes are thought to contain a CuA electron entry site, similar to that of cytochrome c oxidase, and a CuZ catalytic center. The copper anomalous signal was used to solve the crystal structure of N20 reductase from Pseudomonas nautica by multiwavelength anomalous dispersion, to a resolution of 2.4 A. The structure reveals that the CuZ center belongs to a new type of metal cluster, in which four copper ions are liganded by seven histidine residues. N20 binds to this center via a single copper ion. The remaining copper ions might act as an electron reservoir, assuring a fast electron transfer and avoiding the formation of dead-end products.
1072-8368 (Print)1072-8368 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Amino Acid Sequence; Azurin/analogs & derivatives/metabolism; Binding Sites; Catalytic Domain; Copper/*chemistry/*metabolism; Crystallography, X-Ray; Dimerization; Histidine/metabolism; Hydrogen Bonding; Models, Molecular; Molecular Sequence Data; Nitrous Oxide/metabolism; Oxidation-Reduction; Oxidoreductases/*chemistry/*metabolism; Protein Conformation; Protein Folding; Pseudomonas/*enzymology; Structure-Activity Relationship

BiGGER: a new (soft) docking algorithm for predicting protein interactions - Palma, P. N.; Krippahl, L.; Wampler, J. E.; Moura, J. J. (Proteins, 2000)
A new computationally efficient and automated "soft docking" algorithm is described to assist the prediction of the mode of binding between two proteins, using the three-dimensional structures of the unbound molecules. The method is implemented in a software package called BiGGER (Bimolecular Complex Generation with Global Evaluation and Ranking) and works in two sequential steps: first, the complete 6-dimensional binding spaces of both molecules is systematically searched. A population of candidate protein-protein docked geometries is thus generated and selected on the basis of the geometric complementarity and amino acid pairwise affinities between the two molecular surfaces. Most of the conformational changes observed during protein association are treated in an implicit way and test results are equally satisfactory, regardless of starting from the bound or the unbound forms of known structures of the interacting proteins. In contrast to other methods, the entire molecular surfaces are searched during the simulation, using absolutely no additional information regarding the binding sites. In a second step, an interaction scoring function is used to rank the putative docked structures. The function incorporates interaction terms that are thought to be relevant to the stabilization of protein complexes. These include: geometric complementarity of the surfaces, explicit electrostatic interactions, desolvation energy, and pairwise propensities of the amino acid side chains to contact across the molecular interface. The relative functional contribution of each of these interaction terms to the global scoring function has been empirically adjusted through a neural network optimizer using a learning set of 25 protein-protein complexes of known crystallographic structures. In 22 out of 25 protein-protein complexes tested, near-native docked geometries were found with C(alpha) RMS deviations < or =4.0 A from the experimental structures, of which 14 were found within the 20 top ranking solutions. The program works on widely available personal computers and takes 2 to 8 hours of CPU time to run any of the docking tests herein presented. Finally, the value and limitations of the method for the study of macromolecular interactions, not yet revealed by experimental techniques, are discussed.
0887-3585 (Print)0887-3585 (Linking)Journal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.
Keywords: *Algorithms; *Software; Computer Simulation; Humans; Protein Conformation; Protein Folding; Proteins/chemistry/*metabolism

Using cytochrome c(3) to make selenium nanowires - Abdelouas, A.; Gong, W. L.; Lutze, W.; Shelnutt, J. A.; Franco, R.; Moura, I. (Chemistry of Materials, 2000)
We report on a new method to make nanostructures in aqueous solution at room temperature. We used the protein cytochrome c(3) to catalyze reduction of selenate (SeO42-) to selenium Se-0 by dithionite. Reduction was instantaneous. After a week spherical nanoparticles of red Se-0 (about 50 nm diameter) precipitated, followed by self-assembling into crystalline nanowires, typically 1 mu m long. The nanowires were composed of one strand of spherical particles; thicker strands contained several nanoparticles in parallel.
Times Cited: 66

Flavonoids from Ulex species - Maximo, P; Lourenco, A.; Feio, SS; Roseiro, JC (ZEITSCHRIFT FUR NATURFORSCHUNG SECTION C-A JOURNAL OF BIOSCIENCES, 2000)
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Structural model of the Fe-hydrogenase/cytochrome c553 complex combining transverse relaxation-optimized spectroscopy experiments and soft docking calculations - Morelli, X.; Czjzek, M.; Hatchikian, C. E.; Bornet, O.; Fontecilla-Camps, J. C.; Palma, N. P.; Moura, J. J.; Guerlesquin, F. (J Biol Chem, 2000)
Fe-hydrogenase is a 54-kDa iron-sulfur enzyme essential for hydrogen cycling in sulfate-reducing bacteria. The x-ray structure of Desulfovibrio desulfuricans Fe-hydrogenase has recently been solved, but structural information on the recognition of its redox partners is essential to understand the structure-function relationships of the enzyme. In the present work, we have obtained a structural model of the complex of Fe-hydrogenase with its redox partner, the cytochrome c(553), combining docking calculations and NMR experiments. The putative models of the complex demonstrate that the small subunit of the hydrogenase has an important role in the complex formation with the redox partner; 50% of the interacting site on the hydrogenase involves the small subunit. The closest contact between the redox centers is observed between Cys-38, a ligand of the distal cluster of the hydrogenase and Cys-10, a ligand of the heme in the cytochrome. The electron pathway from the distal cluster of the Fe-hydrogenase to the heme of cytochrome c(553) was investigated using the software Greenpath and indicates that the observed cysteine/cysteine contact has an essential role. The spatial arrangement of the residues on the interface of the complex is very similar to that already described in the ferredoxin-cytochrome c(553) complex, which therefore, is a very good model for the interacting domain of the Fe-hydrogenase-cytochrome c(553).
0021-9258 (Print)0021-9258 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Cytochrome c Group/*chemistry; Hydrogenase/*chemistry; Iron-Sulfur Proteins/*chemistry; Magnetic Resonance Spectroscopy/methods; Models, Molecular; Osmolar Concentration

Biochemical/spectroscopic characterization and preliminary X-ray analysis of a new aldehyde oxidoreductase isolated from Desulfovibrio desulfuricans ATCC 27774 - Duarte, R. O.; Archer, M.; Dias, J. M.; Bursakov, S.; Huber, R.; Moura, I.; Romao, M. J.; Moura, J. J. (Biochem Biophys Res Commun, 2000)
Aldehyde oxidoreductase (AOR) activity has been found in different sulfate reducing organisms (Moura, J. J. G., and Barata, B. A. S. (1994) in Methods in Enzymology (Peck, H. D., Jr., and LeGall, J., Eds.), Vol. 243, Chap. 4. Academic Press; Romao, M. J., Knablein, J., Huber, R., and Moura, J. J. G. (1997) Prog. Biophys. Mol. Biol. 68, 121-144). The enzyme was purified to homogeneity from extracts of Desulfovibrio desulfuricans (Dd) ATCC 27774, a sulfate reducer that can use sulfate or nitrate as terminal respiratory substrates. The protein (AORDd) is described as a homodimer (monomer, circa 100 kDa), contains a Mo-MCD pterin, 2 x [2Fe-2S] clusters, and lacks a flavin group. Visible and EPR spectroscopies indicate a close similarity with the AOR purified from Desulfovibrio gigas (Dg) (Barata, B. A. S., LeGall, J., and Moura, J. J. G. (1993) Biochemistry 32, 11559-11568). Activity and substrate specificity for different aldehydes were determined. EPR studies were performed in native and reduced states of the enzyme and after treatment with ethylene glycol and dithiothreitol. The AORDd was crystallized using ammonium sulfate as precipitant and the crystals belong to the space group P6(1)22, with unit cell dimensions a = b = 156.4 and c = 177.1 A. These crystals diffract to beyond 2.5 A resolution and a full data set was measured on a rotating anode generator. The data were used to solve the structure by Patterson Search methods, using the model of AORDg.
0006-291X (Print)0006-291X (Linking)Journal Article
Keywords: *Coenzymes; Aldehyde Oxidoreductases/*chemistry/*isolation & purification/metabolism; Crystallography, X-Ray; Desulfovibrio/*enzymology; Dimerization; Electron Spin Resonance Spectroscopy; Metalloproteins/chemistry; Molecular Weight; Oxidation-Reduction; Protein Structure, Quaternary; Pteridines/chemistry; Spectrophotometry; Substrate Specificity

Crystallization and preliminary X-ray analysis of a membrane-bound nitrite reductase from Desulfovibrio desulfuricans ATCC 27774 - Dias, J. M.; Cunha, C. A.; Teixeira, S.; Almeida, G.; Costa, C.; Lampreia, J.; Moura, J. J.; Moura, I.; Romao, M. J. (Acta Crystallogr D Biol Crystallogr, 2000)
Nitrite reductase from the sulfate-reducing bacterium Desulfovibrio desulfuricans ATCC 27774 is a multihaem (type c) membrane-bound enzyme that catalyzes the dissimilatory conversion of nitrite to ammonia. Crystals of the oxidized form of this enzyme were obtained using PEG and CaCl(2) as precipitants in the presence of 3--(decylmethylammonium)propane-1-sulfonate and belong to the space group P2(1)2(1)2(1), with unit-cell parameters a = 78.94, b = 104.59, c = 143.18 A. A complete data set to 2.30 A resolution was collected using synchrotron radiation at the ESRF. However, the crystals may diffract to beyond 1.7 A and high-resolution data will be collected in the near future.
0907-4449 (Print)0907-4449 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Bacterial Proteins/chemistry/isolation & purification; Crystallization; Desulfovibrio/classification/*enzymology; Electrophoresis, Polyacrylamide Gel; Membrane Proteins/*chemistry/isolation & purification; Nitrite Reductases/*chemistry/isolation & purification; Oxidation-Reduction; Sodium Dodecyl Sulfate; X-ray diffraction

Fisheries Development and Fisheries Dependent Communities in Portugal: Socio-Economic Change and Strategic Planning - António Moniz; Kovács, Ilona; Vicente, Duarte; Ramos, Ana R. (University Library of Munich, Germany, 2000)
The socio-economic subsystem encompassing fisheries may be defined as including not only the harvesting sector but also several related activities occurring both upstream (shipbuilding, gear manufacture) and downstream (processing, distribution and trade). But these closely interrelated economic activities can also be set within a much broader system which would include the ecological, institutional and political influences which frame economic behaviour. The value of this broader conceptualisation is that it treats fisheries not as an isolated and independent economic activity but as part of a more holistic and complex system. This broader perspective is of particular significance when attempting to examine the concept of regional dependence. The socio-economic subsystem for fisheries is dominated by small and medium sized enterprises (SMEs). And Peniche emerges as one of Portugal’s most important fishing ports whether measured in terms of the volume of landings or the total numbers of fishermen. It also has one of the highest levels of fisheries dependence of all coastal municipalities in Portugal with over 20% of its workforce currently engaged in fisheries related employment, faces a daunting and uncertain future. The social fabric of fisheries dependent communities also suffers serious damage; once again, the technocratic approach to management has no solutions to offer. It is essential, therefore, to turn away from the existing approach and to develop instead new forms of intervention; in short, to provide a new vision. This implies change not only to the policy process but also in the attitudes of the social actors and in the preoccupations of fisheries related research. An integrated approach is required based on participative action and the development of an integrated information network.
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Keywords: dependent communities; Fisheries; Peniche; Policy; Portugal; socio-economic system

New quinolizidine alkaloids from Ulex jussiaei - Maximo, P; Lourenco, A. (JOURNAL OF NATURAL PRODUCTS, 2000)
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Trabalho, formação e inovação: Novos problemas para a Sociedade da Informação [Work, training and innovation: new problems for the information society] - António Moniz (University Library of Munich, Germany, 2000)
In this paper for a public debate promoted by the Portuguese President of Republic in 2000, we start from the basic definitions of information society and discuss the main measures proposed as targets in the firld of work and employment. Structural changes are analysed related with the economical and social dimensions of the information society, especially those related to the work organisation, education and training, and the labour markets. Finally recommendations are made on the topics of innovation, employment and development of information society.
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Keywords: education system; Information Society; labour markets; Portugal; training; work organisation

{Fisheries Development and Fisheries Dependent Communities in Portugal: Socio-Economic Change and Strategic Planning} - António Moniz; Kovács, Ilona; Vicente, Duarte; Ramos, Ana R. (University Library of Munich, Germany, 2000)
The socio-economic subsystem encompassing fisheries may be defined as including not only the harvesting sector but also several related activities occurring both upstream (shipbuilding, gear manufacture) and downstream (processing, distribution and trade). But these closely interrelated economic activities can also be set within a much broader system which would include the ecological, institutional and political influences which frame economic behaviour. The value of this broader conceptualisation is that it treats fisheries not as an isolated and independent economic activity but as part of a more holistic and complex system. This broader perspective is of particular significance when attempting to examine the concept of regional dependence. The socio-economic subsystem for fisheries is dominated by small and medium sized enterprises (SMEs). And Peniche emerges as one of Portugal’s most important fishing ports whether measured in terms of the volume of landings or the total numbers of fishermen. It also has one of the highest levels of fisheries dependence of all coastal municipalities in Portugal with over 20% of its workforce currently engaged in fisheries related employment, faces a daunting and uncertain future. The social fabric of fisheries dependent communities also suffers serious damage; once again, the technocratic approach to management has no solutions to offer. It is essential, therefore, to turn away from the existing approach and to develop instead new forms of intervention; in short, to provide a new vision. This implies change not only to the policy process but also in the attitudes of the social actors and in the preoccupations of fisheries related research. An integrated approach is required based on participative action and the development of an integrated information network.
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Keywords: dependent communities; Fisheries; Peniche; Policy; Portugal; socio-economic system

{Trabalho, formação e inovação: Novos problemas para a Sociedade da Informação [Work, training and innovation: new problems for the information society]} - António Moniz (University Library of Munich, Germany, 2000)
In this paper for a public debate promoted by the Portuguese President of Republic in 2000, we start from the basic definitions of information society and discuss the main measures proposed as targets in the firld of work and employment. Structural changes are analysed related with the economical and social dimensions of the information society, especially those related to the work organisation, education and training, and the labour markets. Finally recommendations are made on the topics of innovation, employment and development of information society.
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Keywords: education system; Information Society; labour markets; Portugal; training; work organisation

Evidence for antisymmetric exchange in cuboidal 3Fe-4S (+) clusters - Sanakis, Y.; Macedo, A. L.; Moura, I.; Moura, J. J. G.; Papaefthymiou, V.; Munck, E. (Journal of the American Chemical Society, 2000)
Iron-sulfur clusters with [3Fe-4S] cores are widely distributed in biological systems. In the oxidized state, designated [3Fe-4S](+), these electron-transfer agents have an electronic ground state with S = 1/2, and; they exhibit EPR signals centered at g = 2.01. It has been established by Mossbauer spectroscopy that the three iron sites of the cluster are high-spin Fe3+; and the general properties of the S = 1/2 ground state have been described with the exchange Hamiltonian H-exch = J(12)S(1).S-2 + J(23)S(2).S-3 + J(13)S(1).S-3 Some [3Fe-4S](+) clusters (type 1) have their g-values confined to the range between g = 2.03 and 2.00 while others (type 2) exhibit a continuous distribution of g-values down to g approximate to 1.85. Despite considerable efforts in various laboratories no model has emerged that explains the g-values of type 2 clusters. The 4.2 K spectra of all [3Fe-4S](+) clusters have broad features,which have been simulated in the past by using Fe-57 magnetic hyperfine tensors with anisotropies that are unusually large for high-spin feme sites. It is proposed here that antisymmetric exchange, H-AS = d.(S-1 x S-2 + S-2 x S-3 + S-3 x S-1), is the cause of the g-value shifts in type 2 clusters. We have been able to fit the EPR and Mossbauer spectra of the 3Fe clusters of beef heart aconitase and Desulfovibrio gigas ferredoxin II by using antisymmetric exchange in combination with distributed exchange coupling constants J(12), J(13), and J(23) (J-strain). While antisymmetric exchange is negligible for aconitase (which has a type 1 cluster), fits of the ferredoxin II spectra require \d\ approximate to 0.4 cm(-1). Our studies show that the data of both proteins can lie fit using the same isotropic Fe-57 magnetic hyperfine coupling constant for th three cluster sites, namely a -18.0 MHz for aconitase and a = -18.5 MHz for the D. gigas ferredoxin. The effects of antisymmetric exchange and J-strain on the Mossbauer and EPR spectra are discussed.
Times Cited: 22

Revisiting the catalytic CuZ cluster of nitrous oxide (N2O) reductase. Evidence of a bridging inorganic sulfur - Brown, K.; Djinovic-Carugo, K.; Haltia, T.; Cabrito, I.; Saraste, M.; Moura, J. J.; Moura, I.; Tegoni, M.; Cambillau, C. (J Biol Chem, 2000)
Nitrous-oxide reductases (N2OR) catalyze the two-electron reduction of N(2)O to N(2). The crystal structure of N2ORs from Pseudomonas nautica (Pn) and Paracoccus denitrificans (Pd) were solved at resolutions of 2.4 and 1.6 A, respectively. The Pn N2OR structure revealed that the catalytic CuZ center belongs to a new type of metal cluster in which four copper ions are liganded by seven histidine residues. A bridging oxygen moiety and two other hydroxide ligands were proposed to complete the ligation scheme (Brown, K., Tegoni, M., Prudencio, M., Pereira, A. S., Besson, S., Moura, J. J. G., Moura, I., and Cambillau, C. (2000) Nat. Struct. Biol. 7, 191-195). However, in the CuZ cluster, inorganic sulfur chemical determination and the high resolution structure of Pd N2OR identified a bridging inorganic sulfur instead of an oxygen. This result reconciles the novel CuZ cluster with the hitherto puzzling spectroscopic data.
0021-9258 (Print)0021-9258 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Catalysis; Copper/*chemistry; Electron Spin Resonance Spectroscopy; Oxidoreductases/*chemistry/metabolism; Paracoccus/enzymology; Pseudomonas/enzymology; Sulfur/*chemistry

Desulfoferrodoxin: a modular protein - Ascenso, C.; Rusnak, F.; Cabrito, I.; Lima, M. J.; Naylor, S.; Moura, I.; Moura, J. J. (J Biol Inorg Chem, 2000)
The gene encoding the non-heme iron-containing desulfoferrodoxin from Desulfovibrio vulgaris was cloned in two fragments in order to obtain polypeptides corresponding to the N- and C-terminal domains observed in the tertiary structure. These fragments were expressed in Escherichia coli, purified to homogeneity and biochemically and spectroscopically characterized. Both recombinant fragments behaved as independent metal-binding domains. The N-terminal fragment exhibited properties similar to desulforedoxin, as expected by the presence of a Fe(S-Cys)4 metal binding motif. The C-terminal fragment, which accommodates a Fe(Nepsilon-His)3(Ndelta-His)(S-Cys) center, was shown to have properties similar to neelaredoxin, except for the reaction with superoxide. The activities of desulfoferrodoxin and of the expressed C-terminal fragment were tested with superoxide in the presence and absence of cytochrome c. The results are consistent with superoxide reductase activity and a possible explanation for the low superoxide consumption in the superoxide dismutase activity assays is proposed.
0949-8257 (Print)0949-8257 (Linking)Journal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.
Keywords: Amino Acid Sequence; Base Sequence; Cloning, Molecular; Desulfovibrio vulgaris/genetics; DNA Primers; DNA, Bacterial; Ferredoxins/chemistry/*genetics/metabolism; Molecular Sequence Data; Sequence Homology, Amino Acid; Superoxide Dismutase/metabolism

Redox thermodynamics of low-potential iron-sulfur proteins - Battistuzzi, G.; D'Onofrio, M.; Borsari, M.; Sola, M.; Macedo, A. L.; Moura, J. J.; Rodrigues, P. (J Biol Inorg Chem, 2000)
The enthalpy and entropy changes associated with protein reduction (deltaHdegrees,(rc), deltaSdegrees,(rc)) were determined for a number of low-potential iron-sulfur proteins through variable temperature direct electrochemical experiments. These data add to previous estimates making available, overall, the reduction thermodynamics for twenty species from various sources containing all the different types of metal centers. These parameters are discussed with reference to structural data and calculated electrostatic metal-environment interaction energies, and redox properties of model complexes. This work, which is the first systematic investigation on the reduction thermodynamics of Fe-S proteins, contributes to the comprehension of the determinants of the differences in reduction potential among different protein families within a novel perspective. Moreover, comparison with analogous data obtained previously for electron transport (ET) metalloproteins with positive reduction potentials, i.e., cytochromes c, blue copper proteins, and HiPIPs, helps our understanding of the factors controlling the reduction potential in ET species containing different metal cofactors. The main results of this work can be summarized as follows.
0949-8257 (Print)0949-8257 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Desulfovibrio/chemistry; Electron Transport; Iron-Sulfur Proteins/*chemistry; Oxidation-Reduction; Thermodynamics

A novel protein-bound copper - Molybdenum cluster - George, G. N.; Pickering, I. J.; Yu, E. Y.; Prince, R. C.; Bursakov, S. A.; Gavel, O. Y.; Moura, I.; Moura, J. J. G. (Journal of the American Chemical Society, 2000)
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Times Cited: 48

Purification, characterization, and preliminary crystallographic study of copper-containing nitrous oxide reductase from Pseudomonas nautica 617 - Prudencio, M.; Pereira, A. S.; Tavares, P; Besson, S.; Cabrito, I.; Brown, K.; Samyn, B.; Devreese, B.; Van Beeumen, J.; Rusnak, F.; Fauque, G.; Moura, J. J.; et al. (Biochemistry, 2000)
The aerobic purification of Pseudomonas nautica 617 nitrous oxide reductase yielded two forms of the enzyme exhibiting different chromatographic behaviors. The protein contains six copper atoms per monomer, arranged in two centers named Cu(A) and Cu(Z). Cu(Z) could be neither oxidized nor further reduced under our experimental conditions, and exhibits a 4-line EPR spectrum (g(x)=2.015, A(x)=1.5 mT, g(y)=2.071, A(y)=2 mT, g(z)=2.138, A(z)=7 mT) and a strong absorption at approximately 640 nm. Cu(A) can be stabilized in a reduced EPR-silent state and in an oxidized state with a typical 7-line EPR spectrum (g(x)=g(y)= 2.021, A(x) = A(y)=0 mT, g(z) = 2.178, A(z)= 4 mT) and absorption bands at 480, 540, and approximately 800 nm. The difference between the two purified forms of nitrous oxide reductase is interpreted as a difference in the oxidation state of the Cu(A) center. In form A, Cu(A) is predominantly oxidized (S = (1)/(2), Cu(1.5+)-Cu(1.5+)), while in form B it is mostly in the one-electron reduced state (S = 0, Cu(1+)-Cu(1+)). In both forms, Cu(Z) remains reduced (S = 1/2). Complete crystallographic data at 2.4 A indicate that Cu(A) is a binuclear site (similar to the site found in cytochrome c oxidase) and Cu(Z) is a novel tetracopper cluster [Brown, K., et al. (2000) Nat. Struct. Biol. (in press)]. The complete amino acid sequence of the enzyme was determined and comparisons made with sequences of other nitrous oxide reductases, emphasizing the coordination of the centers. A 10.3 kDa peptide copurified with both forms of nitrous oxide reductase shows strong homology with proteins of the heat-shock GroES chaperonin family.
0006-2960 (Print)0006-2960 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Amino Acid Sequence; Base Sequence; Copper; Crystallography, X-Ray; Molecular Sequence Data; Oxidoreductases/*chemistry/isolation & purification; Pseudomonas/*chemistry/*enzymology; Sequence Alignment

Aldehyde oxidoreductase activity in Desulfovibrio alaskensis NCIMB 13491 EPR assignment of the proximal [2Fe-2S] cluster to the Mo site - Andrade, S. L.; Brondino, C. D.; Feio, M. J.; Moura, I.; Moura, J. J. (Eur J Biochem, 2000)
A novel molybdenum iron-sulfur-containing aldehyde oxidoreductase (AOR) belonging to the xanthine oxidase family was isolated and characterized from the sulfate-reducing bacterium Desulfovibrio alaskensis NCIMB 13491, a strain isolated from a soured oil reservoir in Purdu Bay, Alaska. D. alaskensis AOR is closely related to other AORs isolated from the Desulfovibrio genus. The protein is a 97-kDa homodimer, with 0.6 +/- 0.1 Mo, 3.6 +/- 0.1 Fe and 0.9 +/- 0.1 pterin cytosine dinucleotides per monomer. The enzyme catalyses the oxidation of aldehydes to their carboxylic acid form, following simple Michaelis-Menten kinetics, with the following parameters (for benzaldehyde): K(app/m)= 6.65 microM; V app = 13.12 microM.min(-1); k(app/cat) = 0.96 s(-1). Three different EPR signals were recorded upon long reduction of the protein with excess dithionite: an almost axial signal split by hyperfine interaction with one proton associated with Mo(V) species and two rhombic signals with EPR parameters and relaxation behavior typical of [2Fe-2S] clusters termed Fe/S I and Fe/S II, respectively. EPR results reveal the existence of magnetic interactions between Mo(V) and one of the Fe/S clusters, as well as between the two Fe/S clusters. Redox titration monitored by EPR yielded midpoint redox potentials of -275 and -325 mV for the Fe/S I and Fe/S II, respectively. The redox potential gap between the two clusters is large enough to obtain differentiated populations of these paramagnetic centers. This fact, together with the observed interactions among paramagnetic centers, was used to assign the EPR-distinguishable Fe/S I and Fe/S II to those seen in the reported crystal structures of homologous enzymes.
0014-2956 (Print)0014-2956 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Aldehyde Oxidoreductases/chemistry/isolation & purification/*metabolism; Desulfovibrio/*enzymology; Electron Spin Resonance Spectroscopy; Iron-Sulfur Proteins/*chemistry; Molybdenum/*chemistry; Oxidation-Reduction; Spectrophotometry, Ultraviolet

New Methodological Approaches for Change in Traditional Sectors: The Case of the Portuguese Fisheries Socio-Economic System - António Moniz; Godinho, Manuel M. (University Library of Munich, Germany, 2000)
This paper summarises the methodological approach and main results of the MARHE project (Employment and Human Resources in the Fisheries Socio-Economic System). This project had as its main aim the search for alternative futures for the fisheries sector in Portugal, with particular attention being paid to the human resources situation and the working and living conditions of the fisheries-dependent populations in the coastal areas. This is a particularly interesting case, since fisheries were once an important activity and they are now in deep recession, even though it is generally recognised that the future utilisation of maritime resources offer an immense potential. As part of the research, a Delphi exercise was implemented involving in two successive stages some of the leading actors and experts dealing with the sector in Portugal. Other initiatives were held in the context of the MARHE project providing direct and indirect inputs to the scenarios and recommendations that were put forward in the sequence of the Delphi exercise. Overall the activities described in the paper contributed to the mobilisation of major actors and to discussions that may have practical implication for the future of the sector, if certain conditions are now met in the follow up to the project.
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Keywords: Fisheries; human resources; labour market; Portugal; scenarios

A novel type of catalytic copper cluster in nitrous oxide reductase - Brown, K.; Tegoni, M.; Prudencio, M.; Pereira, A. S.; Besson, S.; Moura, J. J.; Moura, I.; Cambillau, C. (Nature Structural Biology, 2000)
Nitrous oxide (N(2)O) is a greenhouse gas, the third most significant contributor to global warming. As a key process for N(2)O elimination from the biosphere, N(2)O reductases catalyze the two-electron reduction of N(2)O to N(2). These 2 x 65 kDa copper enzymes are thought to contain a CuA electron entry site, similar to that of cytochrome c oxidase, and a CuZ catalytic center. The copper anomalous signal was used to solve the crystal structure of N(2)O reductase from Pseudomonas nautica by multiwavelength anomalous dispersion, to a resolution of 2.4 Angstrom. The structure reveals that the CuZ center belongs to a new type of metal cluster, in which four copper ions are liganded by seven histidine residues. N(2)O binds to this center via a single copper ion. The remaining copper ions might act as an electron reservoir, assuring a fast electron transfer and avoiding the formation of dead-end products.
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{New Methodological Approaches for Change in Traditional Sectors: The Case of the Portuguese Fisheries Socio-Economic System} - António Moniz; Godinho, Manuel M. (University Library of Munich, Germany, 2000)
This paper summarises the methodological approach and main results of the MARHE project (Employment and Human Resources in the Fisheries Socio-Economic System). This project had as its main aim the search for alternative futures for the fisheries sector in Portugal, with particular attention being paid to the human resources situation and the working and living conditions of the fisheries-dependent populations in the coastal areas. This is a particularly interesting case, since fisheries were once an important activity and they are now in deep recession, even though it is generally recognised that the future utilisation of maritime resources offer an immense potential. As part of the research, a Delphi exercise was implemented involving in two successive stages some of the leading actors and experts dealing with the sector in Portugal. Other initiatives were held in the context of the MARHE project providing direct and indirect inputs to the scenarios and recommendations that were put forward in the sequence of the Delphi exercise. Overall the activities described in the paper contributed to the mobilisation of major actors and to discussions that may have practical implication for the future of the sector, if certain conditions are now met in the follow up to the project.
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Keywords: Fisheries; human resources; labour market; Portugal; scenarios

A novel fluorescent chemosensor exhibiting exciplex emission. An example of an elementary molecular machine driven by pH and by light - Bencini, A; Bianchi, A; Lodeiro, C.; Masotti, A.; A. J. Parola; Pina, F; de Melo, JS; Valtancoli, B (Chemical Communications, 2000)
Coordination/detachment of a pendent functionality in the Zn(ii) complex with a macrocyclic ligand L gives rise to on/off switching of exciplex emission, defining an elementary molecular machine whose movements are driven by both pH and light.
ISI Document Delivery No.: 347EQTimes Cited: 50 Cited Reference Count: 21 Bencini, A Bianchi, A Lodeiro, C Masotti, A Parola, AJ Pina, F de Melo, JS Valtancoli, B Masotti, Andrea/D-5306-2009; Pina, Fernando/C-8161-2011; Seixas de Melo, Sergio/A-5388-2008; Lodeiro, Carlos/B-4793-2013; Parola, A. Jorge/F-4048-2010 Masotti, Andrea/0000-0002-2511-5088; Seixas de Melo, Sergio/0000-0001-9708-5079; Lodeiro, Carlos/0000-0001-5582-5446; Parola, A. Jorge/0000-0002-1333-9076 Royal soc chemistry Cambridge

Adaptive Regulation of Arc Welding Temperature with Parallel Integral Action - Santos, TO; Caetano, RB; Lemos, J. M.; Coito, F.J. (2000)

Characterization of the murMN operon involved in the synthesis of branched peptidoglycan peptides in Streptococcus pneumoniae - S. R. Filipe; M. G. Pinho; A. Tomasz (J. Biol. Chem, 2000)

Distribution of the mosaic structured murM genes among natural populations of Streptococcus pneumoniae - S. R. Filipe; E. Severina; A. Tomasz (J. Bacteriol, 2000)

Inhibition of the expression of penicillin resistance in Streptococcus pneumoniae by inactivation of cell wall muropeptide branching genes - S. R. Filipe; A. Tomasz (Proc. Natl. Acad. Sci. USA, 2000)

Relaxation Studies in PEO/PMMA Blends - M. Dionísio; A. C. Fernandes; J.F. Mano; N. T. Correia; R. C. Sousa (Macromolecules, 2000)

A Surface Chemistry Experiment Using an Inexpensive W Contact Angle Goniometer - M. Dionísio; J. Sotomayor (Journal of Chemical Education, 2000)

Adaptive Regulation of Arc Welding Temperature with Parallel Integral Action - Santos, TO; Caetano, RB; Lemos, J. M.; Coito, F.J. (2000)
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Anomalous fluorescence of linear poly(methylphenylsiloxane) in dilute solution at temperatures below-50 degrees C - Dias, FB; J. C. Lima; Macanita, AL; Clarson, SJ; Horta, A; Pierola, IF (Macromolecules, 2000)
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Artificial chemical systems capable of mimicking some elementary properties of neurons - Pina, F; Melo, MJ; Maestri, M; Passaniti, P.; Balzani, V. (Journal of the American Chemical Society, 2000)
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Times Cited: 135 Melo, Maria Joao/C-8594-2011; Pina, Fernando/C-8161-2011; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Melo, Maria Joao/0000-0001-7393-6801; Pina, Fernando/0000-0001-8529-6848; 136

Biochemical/spectroscopic characterization and preliminary X-ray analysis of a new aldehyde oxidoreductase isolated from Desulfovibrio desulfuricans ATCC 27774 - Duarte, R. O.; Archer, M.; Dias, J. M.; Bursakov, S.; Huber, R.; Moura, I.; Romao, M. J.; Moura, J. J. G. (Biochemical and Biophysical Research Communications, 2000)
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Times Cited: 8 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 8

Conditions Of Inter-Firm Co-Operation In A Virtual Enterprise Concept : The Case Of Automotive Sector In Portugal - António Moniz; Kovács, Ilona (University Library of Munich, Germany, 2000)
One can assist to significant changes in the organisation of manufacturing systems during the last years. Lean production, network enterprise or the virtual enterprises are reference concepts of the re-organisation of manufacturing systems. Some authors mention a new enterprise paradigm, of generalisation of intelligent manufacture, organised in networks and assisted by information and communication technologies. The first part of the paper develops a critical approach to the illusion connected to these concepts, calling the attention to the diversity of the type of relationships among firms. If virtual enterprises (VE) are networks of firms with intensive usage of ICT, one can verify a predominance of a technicist perspective. This one considers that the development of VEs is a technological problem, of development and management of information systems, and of entrepreneurial share of different databases. Sociology can be useful, even fundamental in an anthropocentric approach. The last part of the paper is on the Portuguese situation in the automobile sector, approaching the types of entrepreneurial organisation.
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Keywords: automobile sector; manufacturing systems; networks; Portugal; virtual enterprises

Crystallization and preliminary X-ray analysis of a membrane-bound nitrite reductase from Desulfovibrio desulfuricans ATCC 27774 - Dias, J. M.; Cunha, C. A.; Teixeira, S.; Almeida, G.; Costa, C.; Lampreia, J.; Moura, J. J. G.; Moura, I.; Romao, M. J. (Acta Crystallographica Section D-Biological Crystallography, 2000)
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Times Cited: 6 Costa, Cristina/A-9966-2012; Romao, Maria/A-4115-2013; Lampreia, Jorge/A-9927-2012; Almeida, Maria Gabriela/D-8265-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Costa, Cristina/0000-0002-8611-9023; Romao, Maria/0000-0002-3004-0543; Lampreia, Jorge/0000-0001-8846-0041; Almeida, Maria Gabriela/0000-0002-4508-7379; Moura, Isabel/0000-0003-0971-4977; 2 6

Dynamics of cyclic methylphenyltrisiloxane in the picosecond to nanosecond time range - Dias, FB; J. C. Lima; Macanita, AL; Horta, A; Pierola, IF (Journal of Physical Chemistry a, 2000)
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Fluorescent chemosensors containing polyamine receptors - Pina, F; Bernardo, M. A.; Garcia-Espana, E (European Journal of Inorganic Chemistry, 2000)
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Times Cited: 94 Pina, Fernando/C-8161-2011; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Pina, Fernando/0000-0001-8529-6848; 95

Gene sequence and crystal structure of the aldehyde oxidoreductase from Desulfovibrio desulfuricans ATCC 27774 - Rebelo, J.; Macieira, S.; Dias, J. M.; Huber, R.; Ascenso, C. S.; Rusnak, F.; Moura, J. J. G.; Moura, I.; Romao, M. J. (Journal of Molecular Biology, 2000)
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Times Cited: 46 Romao, Maria/A-4115-2013; Moura, Jose/D-6426-2013; REQUIMTE, SMB/M-5694-2013; Moura, Isabel/D-6339-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; Moura, Isabel/0000-0003-0971-4977; 46

Ionic strength dependence of the non-physiological electron transfer between flavodoxin and cytochrome c(553) from D-vulgaris - Sadeghi, S. J.; Valetti, F.; Cunha, C. A.; Romao, M. J.; Soares, C.M.; Gilardi, G. (Journal of Biological Inorganic Chemistry, 2000)
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Times Cited: 8 GILARDI, GIANFRANCO/B-4523-2010; Soares, Claudio/E-2675-2012; Romao, Maria/A-4115-2013; REQUIMTE, SMB/M-5694-2013; REQUIMTE, UCIBIO/N-9846-2013 Romao, Maria/0000-0002-3004-0543; 8

Light and pH switching between the various forms of the 4 '-methylflavylium cation - Maestri, M; Pina, F; Roque, A.; Passaniti, P. (Journal of Photochemistry and Photobiology a-Chemistry, 2000)
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Times Cited: 8 Pina, Fernando/C-8161-2011; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Pina, Fernando/0000-0001-8529-6848; 8

Light scattering study of water-in-oil AOT microemulsions with poly(oxy)ethylene - Laia, C. A. T.; Brown, W.; Almgren, M.; Costa, S. M. B. (Langmuir, 2000)
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Times Cited: 16 Laia, Cesar/C-8651-2011; Costa, Silvia/H-7766-2012; REQUIMTE, PHOTO/M-4229-2013; Costa, Silvia/M-8085-2013; REQUIMTE, LAQV/N-9835-2013 Laia, Cesar/0000-0001-6410-6072; Costa, Silvia/0000-0002-6978-8903; 16

Multipredictive adaptive control of arc welding trailing centerline temperature - Santos, TO; Caetano, RB; Lemos, J. M.; Coito, F.J. (Control Systems Technology, IEEE Transactions on, 2000)
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On the red colour of raspberry (Rubus idaeus) - Melo, MJ; Moncada, MC; Pina, F (Tetrahedron Letters, 2000)
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Times Cited: 9 Melo, Maria Joao/C-8594-2011; Pina, Fernando/C-8161-2011; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Melo, Maria Joao/0000-0001-7393-6801; Pina, Fernando/0000-0001-8529-6848; 11

pH modulation of the luminescence emission of a new europium cryptate complex - Bazzicalupi, C; Bencini, A; Bianchi, A; Giorgi, C; Fusi, V.; Masotti, A.; Valtancoli, B; Roque, A.; Pina, F (Chemical Communications, 2000)
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Times Cited: 62 Masotti, Andrea/D-5306-2009; Pina, Fernando/C-8161-2011; Fusi, Vieri/H-1006-2012; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Masotti, Andrea/0000-0002-2511-5088; Pina, Fernando/0000-0001-8529-6848; 63

Photochemistry of luteolinidin - "Write-lock-read-unlock-erase'' with a natural compound - Melo, MJ; Moura, S.; Roque, A.; Maestri, M; Pina, F (Journal of Photochemistry and Photobiology a-Chemistry, 2000)
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Times Cited: 11 Melo, Maria Joao/C-8594-2011; Pina, Fernando/C-8161-2011; REQUIMTE, PHOTO/M-4229-2013; REQUIMTE, LAQV/N-9835-2013 Melo, Maria Joao/0000-0001-7393-6801; Pina, Fernando/0000-0001-8529-6848; 11

Photophysical properties of hydroxy-substituted flavothiones - Elisei, F; J. C. Lima; Ortica, F; Aloisi, GG; Costa, M; Leitao, E; Abreu, I; Dias, A; Bonifacio, V; Medeiros, J; Macanita, AL; Becker, RS (Journal of Physical Chemistry a, 2000)
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Purification, characterization, and preliminary crystallographic study of copper-containing nitrous oxide reductase from Pseudomonas nautica 617 - Prudencio, M.; Pereira, A. S.; Tavares, P; Besson, S.; Cabrito, I.; Brown, K.; Samyn, B.; Devreese, B.; Van Beeumen, J.; Rusnak, F.; Fauque, G.; Moura, J. J. G.; et al. (Biochemistry, 2000)
The aerobic purification of Pseudomonas nautica 617 nitrous oxide reductase yielded two forms of the enzyme exhibiting different chromatographic behaviors. The protein contains six copper atoms per monomer, arranged in two centers named CUA and Cut. Cut could be neither oxidized nor further reduced under our experimental conditions, and exhibits a 4-line EPR spectrum (g(x)= 2.015, A(x) = 1.5 mT, g(y) = 2.071, A(y) = 2 mT, g(z) = 2.138, A(z) = 7 mT) and a strong absorption at similar to 640 nm. Cu-A can be stabilized in a reduced EPR-silent state and in an oxidized state with a typical 7-line EPR spectrum (g(x) g(y) = 2.021, A(x) = A(y) = 0 T, g(z) =0.178, A(z) = 4 mT) and absorption bands at 480, 540, and similar to 800 nm. The difference between the two purified forms of nitrous oxide reductase is interpreted as a difference in the oxidation state of the CuA center. In form A, CUA is predominantly oxidized (S = 1/2, Cu1.5+-Cu1.5+), while in form B it is mostly in the one-electron reduced state (S = 0, Cu1+-Cu1+). In both forms, Cu-Z remains reduced (S = 1/2). Complete crystallographic data at 2.4 Angstrom indicate that Cu-A is a binuclear site (similar to the site found in cytochrome c oxidase) and Cu-Z is a novel tetracopper cluster [Brown, K., et ai. (2000) Nat. Struct. Biol. (in press)]. The complete amino acid sequence of the enzyme was determined and comparisons made with sequences of other nitrous oxide reductases, emphasizing the coordination of the centers. A 10.3 kDa peptide copurified with both forms of nitrous oxide reductase shows strong homology with proteins of the heat-shock GroES chaperonin family.
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Redox potential measurements of the Mycobacterium tuberculosis heme protein KatG and the isoniazid-resistant enzyme KatG(S315T): Insights into isoniazid activation - Wengenack, N. L.; Lopes, H.; Kennedy, M. J.; Tavares, P; Pereira, A. S.; Moura, I.; Moura, J. J. G.; Rusnak, F. (Biochemistry, 2000)
Mycobacterium tuberculosis KatG is a multifunctional heme enzyme responsible for activation of the antibiotic isoniazid. A KatG(S315T) point mutation is found in >50% of isoniazid-resistant clinical isolates. Since isoniazid activation is thought to involve an oxidation reaction, the redox potential of KatG was determined using cyclic voltammetry, square wave voltammetry, and spectroelectrochemical titrations. Isoniazid activation may proceed via a cytochrome P450-like mechanism. Therefore, the possibility that substrate binding by KatG leads to an increase in the heme redox potential and the possibility that KatG(S315T) confers isoniazid resistance by altering the redox potential were examined. Effects of the heme spin state on the reduction potentials of KatG and KatG(S315T) were also determined. Assessment of the Fe3+/Fe2+ couple gave a midpoint potential of ca. -50 mV for both KatG and KatG(S315T). In contrast to cytochrome P450s, addition of substrate had no significant effect on either the KatG or KatG(S315T) redox potential. Conversion of the heme to a low-spin configuration resulted in a -150 to -200 mV shift of the KatG and KatG(S315T) redox potentials. These results suggest that isoniazid resistance conferred by KatG(S315T) is not mediated through changes in the heme redox potential. The redox potentials of isoniazid were also determined using cyclic and square wave voltammetry, and the results provide evidence that the ferric KatG and KatG(S315T) midpoint potentials are too low to promote isoniazid oxidation without formation of a high-valent enzyme intermediate such as compounds I and IT or oxyferrous KatG.
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Temperature and composition dependence of the structure of isooctane/AOT microemulsion L-2 phases with glycerol and formamide: A light scattering study - Laia, C. A. T.; Brown, W.; Almgren, M.; Costa, S. M. B. (Langmuir, 2000)
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Times Cited: 20 Laia, Cesar/C-8651-2011; Costa, Silvia/H-7766-2012; REQUIMTE, PHOTO/M-4229-2013; Costa, Silvia/M-8085-2013; REQUIMTE, LAQV/N-9835-2013 Laia, Cesar/0000-0001-6410-6072; Costa, Silvia/0000-0002-6978-8903; 20

{Conditions Of Inter-Firm Co-Operation In A Virtual Enterprise Concept : The Case Of Automotive Sector In Portugal} - António Moniz; Kovács, Ilona (University Library of Munich, Germany, 2000)
One can assist to significant changes in the organisation of manufacturing systems during the last years. Lean production, network enterprise or the virtual enterprises are reference concepts of the re-organisation of manufacturing systems. Some authors mention a new enterprise paradigm, of generalisation of intelligent manufacture, organised in networks and assisted by information and communication technologies. The first part of the paper develops a critical approach to the illusion connected to these concepts, calling the attention to the diversity of the type of relationships among firms. If virtual enterprises (VE) are networks of firms with intensive usage of ICT, one can verify a predominance of a technicist perspective. This one considers that the development of VEs is a technological problem, of development and management of information systems, and of entrepreneurial share of different databases. Sociology can be useful, even fundamental in an anthropocentric approach. The last part of the paper is on the Portuguese situation in the automobile sector, approaching the types of entrepreneurial organisation.
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Keywords: automobile sector; manufacturing systems; networks; Portugal; virtual enterprises

A cytochrome c peroxidase from Pseudomonas nautica 617 active at high ionic strength: expression, purification and characterization - Alves, T.; Besson, S.; Duarte, L. C.; Pettigrew, G. W.; Girio, F. M. F.; Devreese, B.; Vandenberghe, I.; Van Beeumen, J.; Fauque, G.; Moura, I. (Biochimica Et Biophysica Acta-Protein Structure and Molecular Enzymology, 1999)
Cytochrome c peroxidase was expressed in cells of Pseudomonas nautica strain 617 grown under microaerophilic conditions. The 36.5 kDa dihaemic enzyme was purified to electrophoretic homogeneity in three chromatographic steps. N-terminal sequence comparison showed that the Ps. nautica enzyme exhibits a high similarity with the corresponding proteins from Paracoccus denitrificans and Pseudomonas aeruginosa. UV-visible spectra confirm calcium activation of the enzyme through spin state transition of the peroxidatic haem. Monohaemic cytochrome c(552) from Ps. nautica was identified as the physiological electron donor, with a half-saturating concentration of 122 mu M and allowing a maximal catalytic centre activity of 116 000 min(-1). Using this cytochrome the enzyme retained the same activity even at high ionic strength. There are indications that the interactions between the two redox partners are mainly hydrophobic in nature. (C) 1999 Elsevier Science B.V. All rights reserved.
Times Cited: 27

A Diels-Alder, retro-Diels-Alder approach to arcyriaflavin-A - Marques, M M B; A. M. Lobo; Prabhakar, S.; Branco, P. S. (TETRAHEDRON LETTERS, 1999)
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Observation of ligand-based redox chemistry at the active site of a molybdenum enzyme - George, G. N.; Costa, C.; Moura, J. J. G.; Moura, I. (Journal of the American Chemical Society, 1999)
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Times Cited: 32

Reactions of 2-acylthiazolium salts with N-arylhydroxylamines - Ferreira, LM; A. M. Lobo; Prabhakar, S.; Teixeira, AC (TETRAHEDRON, 1999)
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Cross-linking between cytochrome c3 and flavodoxin from Desulfovibrio gigas - Correia, C.; Monzani, E.; Moura, I.; Lampreia, J.; Moura, J. J. (Biochem Biophys Res Commun, 1999)
Tetraheme cytochrome c3 (13 kDa) and flavodoxin (16 kDa), are small electron transfer proteins that have been used to mimic, in vitro, part of the electron-transfer chain that operates between substract electron donors and respiratory electron acceptors partners in Desulfovibrio species (Palma, N., Moura, I., LeGall, J., Van Beeumen, J., Wampler, J., Moura, J. J. G. (1994) Biochemistry 33, 6394-6407). The electron transfer between these two proteins is believed to occur through the formation of a specific complex where electrostatic interaction is the main driving force (Stewart, D., LeGall, J., Moura, I., Moura, J.J.G., Peck, H.D., Xavier, A.V., Weiner, P.K. and Wampler, J.E. (1988) Biochemistry 27, 2444-2450, Stewart, D., LeGall, J., Moura, I., Moura, J.J.G., Peck, H.D., Xavier, A.V., Weiner, P., Wampler, J. (1989) Eur. J. Biochem. 185, 695-700). In order to obtain structural information of the pre-complex, a covalent complex between the two proteins was prepared. A water-soluble carbodiimide [EDC (1-ethyl-3(3 dimethylaminopropyl) carbodiimide hydrochloride] was used for the cross linking reaction. The reaction was optimized varying a wide number of experimental parameters such as ionic strength, protein and cross linker concentration, and utilization of different cross linkers and reaction time between the crosslinker and proteins.
0006-291X (Print)0006-291X (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: *Cross-Linking Reagents; Chromatography, High Pressure Liquid; CME-Carbodiimide/analogs & derivatives; Cytochrome c Group/chemistry/*metabolism; Desulfovibrio/*metabolism; Dose-Response Relationship, Drug; Electron Transport; Ethyldimethylaminopropyl Carbodiimide; Flavodoxin/chemistry/*metabolism; Hydrogen-Ion Concentration; Molecular Weight; Osmolar Concentration; Protein Binding/drug effects; Sodium Chloride/pharmacology; Static Electricity; Succinimides; Time Factors

Biochemical and spectroscopic characterization of overexpressed fuscoredoxin from Escherichia coli - Pereira, A. S.; Tavares, P; Krebs, C.; Huynh, B. H.; Rusnak, F.; Moura, I.; Moura, J. J. (Biochem Biophys Res Commun, 1999)
Fuscoredoxin is a unique iron containing protein of yet unknown function originally discovered in the sulfate reducers of the genus Desulfovibrio. It contains two iron-sulfur clusters: a cubane [4Fe-4S] and a mixed oxo- and sulfido-bridged 4Fe cluster of unprecedented structure. The recent determination of the genomic sequence of Escherichia coli (E. coli) has revealed a homologue of fuscoredoxin in this facultative microbe. The presence of this gene in E. coli raises interesting questions regarding the function of fuscoredoxin and whether this gene represents a structural homologue of the better-characterized Desulfovibrio proteins. In order to explore the latter, an overexpression system for the E. coli fuscoredoxin gene was devised. The gene was cloned from genomic DNA by use of the polymerase chain reaction into the expression vector pT7-7 and overexpressed in E. coli BL21(DE3) cells. After two chromatographic steps a good yield of recombinant protein was obtained (approximately 4 mg of pure protein per liter of culture). The purified protein exhibits an optical spectrum characteristic of the homologue from D. desulfuricans, indicating that cofactor assembly was accomplished. Iron analysis indicated that the protein contains circa 8 iron atoms/molecule which were shown by EPR and Mossbauer spectroscopies to be present as two multinuclear clusters, albeit with slightly altered spectroscopic features. A comparison of the primary sequences of fuscoredoxins is presented and differences on cluster coordination modes are discussed on the light of the spectroscopic data.
0006-291X (Print)0006-291X (Linking)Journal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.
Keywords: Amino Acid Sequence; Bacterial Proteins; Escherichia coli/*chemistry; Iron-Sulfur Proteins/*chemistry/*isolation & purification/*metabolism; Molecular Sequence Data; Recombinant Proteins/chemistry; Sequence Homology, Amino Acid; Spectroscopy, Mossbauer

MAD structure of Pseudomonas nautica dimeric cytochrome c552 mimicks the c4 Dihemic cytochrome domain association - Brown, K.; Nurizzo, D.; Besson, S.; Shepard, W.; Moura, J.; Moura, I.; Tegoni, M.; Cambillau, C. (J Mol Biol, 1999)
The monohemic cytochrome c552from Pseudomonas nautica (c552-Pn) is thought to be the electron donor to cytochrome cd1, the so-called nitrite reductase (NiR). It shows as high levels of activity and affinity for the P. nautica NiR (NiR-Pn), as the Pseudomonas aeruginosa enzyme (NiR-Pa). Since cytochrome c552is by far the most abundant electron carrier in the periplasm, it is probably involved in numerous other reactions. Its sequence is related to that of the c type cytochromes, but resembles that of the dihemic c4cytochromes even more closely. The three-dimensional structure of P. nautica cytochrome c552has been solved to 2.2 A resolution using the multiple wavelength anomalous dispersion (MAD) technique, taking advantage of the presence of the eight Fe heme ions in the asymmetric unit. Density modification procedures involving 4-fold non-crystallographic averaging yielded a model with an R -factor value of 17.8 % (Rfree=20.8 %). Cytochrome c552forms a tight dimer in the crystal, and the dimer interface area amounts to 19% of the total cytochrome surface area. Four tighly packed dimers form the eight molecules of the asymmetric unit. The c552dimer is superimposable on each domain of the monomeric cytochrome c4from Pseudomomas stutzeri (c4-Ps), a dihemic cytochrome, and on the dihemic c domain of flavocytochrome c of Chromatium vinosum (Fcd-Cv). The interacting residues which form the dimer are both similar in character and position, which is also true for the propionates. The dimer observed in the crystal also exists in solution. It has been hypothesised that the dihemic c4-Ps may have evolved via monohemic cytochrome c gene duplication followed by evolutionary divergence and the adjunction of a connecting linker. In this process, our dimeric c552structure might be said to constitute a "living fossile" occurring in the course of evolution between the formation of the dimer and the gene duplication and fusion. The availability of the structure of the cytochrome c552-Pn and that of NiR from P. aeruginosa made it possible to identify putative surface patches at which the docking of c552to NiR-Pn may occur.
0022-2836 (Print)0022-2836 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Amino Acid Sequence; Cytochrome c Group/*chemistry/metabolism; Dimerization; Microspectrophotometry; Models, Molecular; Molecular Mimicry; Molecular Sequence Data; Nitrite Reductases/metabolism; Protein Conformation; Pseudomonas/*chemistry; Sequence Homology, Amino Acid; Solutions

Improving Distributed Intelligence in Complex Innovation Systems - Kuhlmann, Stefan; Boekholt, Patries; Georghiou, Luke; Guy, Ken; Heraud, Jean-Alain; Laredo, Philippe; Lemola, Tarmo; Loveridge, Denis; Luukkonen, Terttu; António Moniz; Polt, Wolfgang; Ri (University Library of Munich, Germany, 1999)
Science and technology (S&T) are considered to be a central source, or at least a basic medium, of societal and industrial innovation, while innovation is conceived to basically feed the regeneration of our welfare. The suppliers of S&T in Europe as well as the users of their „products“, are confronted with a number of challenges today. We want to stress here that it was not the primary goal of our Advanced Science & Technology Policy Planning (ASTPP) Network to come up with proposals how the strategic character of European S&T policies could be strengthened. The ASTPP-network instead focuses on one aspect: the provision of strategic intelligence necessary to identify and develop strategic choices. The underlying hypothesis is that the existing body of experiences with technology foresight, technology assessment and S/T policy evaluation provides a basis for the development of an advanced S&T policy „planning“ approach by trying to enhance, interlink or even integrate the growing, but still dispersed experience in these three areas of intelligence. By „intelligent“ we mean that the inter-relatedness of S&T, industrial efforts, societal needs and political interventions becomes more transparent so that interactive collaboration between them will be facilitated.
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Keywords: assessment; evaluation; innovation; technology foresight; technology policy

{Improving Distributed Intelligence in Complex Innovation Systems} - Kuhlmann, Stefan; Boekholt, Patries; Georghiou, Luke; Guy, Ken; Heraud, Jean-Alain; Laredo, Philippe; Lemola, Tarmo; Loveridge, Denis; Luukkonen, Terttu; António Moniz; Polt, Wolfgang; Rip (University Library of Munich, Germany, 1999)
Science and technology (S&T) are considered to be a central source, or at least a basic medium, of societal and industrial innovation, while innovation is conceived to basically feed the regeneration of our welfare. The suppliers of S&T in Europe as well as the users of their „products“, are confronted with a number of challenges today. We want to stress here that it was not the primary goal of our Advanced Science & Technology Policy Planning (ASTPP) Network to come up with proposals how the strategic character of European S&T policies could be strengthened. The ASTPP-network instead focuses on one aspect: the provision of strategic intelligence necessary to identify and develop strategic choices. The underlying hypothesis is that the existing body of experiences with technology foresight, technology assessment and S/T policy evaluation provides a basis for the development of an advanced S&T policy „planning“ approach by trying to enhance, interlink or even integrate the growing, but still dispersed experience in these three areas of intelligence. By „intelligent“ we mean that the inter-relatedness of S&T, industrial efforts, societal needs and political interventions becomes more transparent so that interactive collaboration between them will be facilitated.
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Keywords: assessment; evaluation; innovation; technology foresight; technology policy

Structural studies by X-ray diffraction on metal substituted desulforedoxin, a rubredoxin-type protein - Archer, M.; Carvalho, A. L.; Teixeira, S.; Moura, I.; Moura, J. J.; Rusnak, F.; Romao, M. J. (Protein Sci, 1999)
Desulforedoxin (Dx), isolated from the sulfate reducing bacterium Desulfovibrio gigas, is a small homodimeric (2 x 36 amino acids) protein. Each subunit contains a high-spin iron atom tetrahedrally bound to four cysteinyl sulfur atoms, a metal center similar to that found in rubredoxin (Rd) type proteins. The simplicity of the active center in Dx and the possibility of replacing the iron by other metals make this protein an attractive case for the crystallographic analysis of metal-substituted derivatives. This study extends the relevance of Dx to the bioinorganic chemistry field and is important to obtain model compounds that can mimic the four sulfur coordination of metals in biology. Metal replacement experiments were carried out by reconstituting the apoprotein with In3+, Ga3+, Cd2+, Hg2+, and Ni2+ salts. The In3+ and Ga3+ derivatives are isomorphous with the iron native protein; whereas Cd2+, Hg2+, and Ni2+ substituted Dx crystallized under different experimental conditions, yielding two additional crystal morphologies; their structures were determined by the molecular replacement method. A comparison of the three-dimensional structures for all metal derivatives shows that the overall secondary and tertiary structures are maintained, while some differences in metal coordination geometry occur, namely, bond lengths and angles of the metal with the sulfur ligands. These data are discussed in terms of the entatic state theory.
0961-8368 (Print)0961-8368 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Amino Acid Sequence; Bacterial Proteins/*chemistry; Crystallography, X-Ray; Iron-Sulfur Proteins/*chemistry; Metals/*chemistry; Models, Molecular; Molecular Sequence Data; Protein Conformation; Recombinant Proteins/chemistry; Rubredoxins/chemistry; Sequence Homology, Amino Acid

Crystal structure of the first dissimilatory nitrate reductase at 1.9 A solved by MAD methods - Dias, J. M.; Than, M. E.; Humm, A.; Huber, R.; Bourenkov, G. P.; Bartunik, H. D.; Bursakov, S.; Calvete, J.; Caldeira, J.; Carneiro, C.; Moura, J. J.; Moura, I.; et al. (Structure, 1999)
BACKGROUND: The periplasmic nitrate reductase (NAP) from the sulphate reducing bacterium Desulfovibrio desulfuricans ATCC 27774 is induced by growth on nitrate and catalyses the reduction of nitrate to nitrite for respiration. NAP is a molybdenum-containing enzyme with one bis-molybdopterin guanine dinucleotide (MGD) cofactor and one [4Fe-4S] cluster in a single polypeptide chain of 723 amino acid residues. To date, there is no crystal structure of a nitrate reductase. RESULTS: The first crystal structure of a dissimilatory (respiratory) nitrate reductase was determined at 1.9 A resolution by multiwavelength anomalous diffraction (MAD) methods. The structure is folded into four domains with an alpha/beta-type topology and all four domains are involved in cofactor binding. The [4Fe-4S] centre is located near the periphery of the molecule, whereas the MGD cofactor extends across the interior of the molecule interacting with residues from all four domains. The molybdenum atom is located at the bottom of a 15 A deep crevice, and is positioned 12 A from the [4Fe-4S] cluster. The structure of NAP reveals the details of the catalytic molybdenum site, which is coordinated to two MGD cofactors, Cys140, and a water/hydroxo ligand. A facile electron-transfer pathway through bonds connects the molybdenum and the [4Fe-4S] cluster. CONCLUSIONS: The polypeptide fold of NAP and the arrangement of the cofactors is related to that of Escherichia coli formate dehydrogenase (FDH) and distantly resembles dimethylsulphoxide reductase. The close structural homology of NAP and FDH shows how small changes in the vicinity of the molybdenum catalytic site are sufficient for the substrate specificity.
0969-2126 (Print)0969-2126 (Linking)Comparative StudyJournal ArticleResearch Support, Non-U.S. Gov't
Keywords: Amino Acid Sequence; Cloning, Molecular; Crystallography, X-Ray; Desulfovibrio/*enzymology; Guanine Nucleotides/chemistry/metabolism; Iron-Sulfur Proteins/chemistry/metabolism; Models, Molecular; Molecular Sequence Data; Molybdenum/metabolism; Nitrate Reductase; Nitrate Reductases/*chemistry/isolation & purification/metabolism; Organometallic Compounds/chemistry/metabolism; Polymerase Chain Reaction; Protein Conformation; Protein Structure, Secondary; Recombinant Proteins/chemistry/isolation & purification/metabolism; Sequence Alignment; Sequence Homology, Amino Acid; X-ray diffraction

Novel acid catalysed 1,4-addition-type ring-opening polymerisation of cyclic phosphorimidates - E. J. Cabrita; Candeias, SX; A. M. Ramos; C. A. M. Afonso; Santos, A. G. (Tetrahedron Letters, 1999)
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Purification and characterization of a tungsten-containing formate dehydrogenase from Desulfovibrio gigas - Almendra, M. J.; Brondino, C. D.; Gavel, O.; Pereira, A. S.; Tavares, P; Bursakov, S.; Duarte, R.; Caldeira, J.; Moura, J. J.; Moura, I. (Biochemistry, 1999)
An air-stable formate dehydrogenase (FDH), an enzyme that catalyzes the oxidation of formate to carbon dioxide, was purified from the sulfate reducing organism Desulfovibrio gigas (D. gigas) NCIB 9332. D. gigas FDH is a heterodimeric protein [alpha (92 kDa) and beta (29 kDa) subunits] and contains 7 +/- 1 Fe/protein and 0.9 +/- 0.1 W/protein. Selenium was not detected. The UV/visible absorption spectrum of D. gigas FDH is typical of an iron-sulfur protein. Analysis of pterin nucleotides yielded a content of 1.3 +/- 0.1 guanine monophosphate/mol of enzyme, which suggests a tungsten coordination with two molybdopterin guanine dinucleotide cofactors. Both Mossbauer spectroscopy performed on D. gigas FDH grown in a medium enriched with (57)Fe and EPR studies performed in the native and fully reduced state of the protein confirmed the presence of two [4Fe-4S] clusters. Variable-temperature EPR studies showed the presence of two signals compatible with an atom in a d(1) configuration albeit with an unusual relaxation behavior as compared to the one generally observed for W(V) ions.
0006-2960 (Print)0006-2960 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Aerobiosis; Chromatography, Gel; Desulfovibrio/*enzymology; Electron Spin Resonance Spectroscopy; Formate Dehydrogenases/*chemistry/*isolation & purification/metabolism; Guanine Nucleotides/analysis; Metalloproteins/chemistry/isolation & purification/metabolism; Molecular Weight; Pterins/analysis; Spectrophotometry, Ultraviolet; Spectroscopy, Mossbauer; Tungsten/*chemistry

New syntheses of DNA adducts from methylated anilines present in tobacco smoke - Branco, P. S.; A.M.M. Antunes; M.M. Marques; Chiarelli, MP; A. M. Lobo; Prabhakar, S. (CHEMICAL RESEARCH IN TOXICOLOGY, 1999)
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The solution structure of a [3Fe-4S] ferredoxin: oxidised ferredoxin II from Desulfovibrio gigas - Goodfellow, B. J.; Macedo, A. L.; Rodrigues, P.; Moura, I.; Wray, V.; Moura, J. J. (J Biol Inorg Chem, 1999)
The use of standard 2D NMR experiments in combination with 1D NOE experiments allowed the assignment of 51 of the 58 spin systems of oxidised [3Fe4S] ferredoxin isolated from Desulfovibrio gigas. The NMR solution structure was determined using data from 1D NOE and 2D NOESY spectra, as distance constraints, and information from the X-ray structure for the spin systems not detected by NMR in torsion angle dynamics calculations to produce a family of 15 low target function structures. The quality of the NMR family, as judged by the backbone r.m.s.d. values, was good (0.80 A), with the majority of phi/psi angles falling within the allowed region of the Ramachandran plot. A comparison with the X-ray structure indicated that the overall global fold is very similar in solution and in the solid state. The determination of the solution structure of ferredoxin II (FdII) in the oxidised state (FdIIox) opens the way for the determination of the solution structure of the redox intermediate state of FdII (FdII(int)), for which no X-ray structure is available.
0949-8257 (Print)0949-8257 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Amino Acid Sequence; Desulfovibrio/*chemistry; Ferredoxins/*chemistry/*metabolism; Magnetic Resonance Spectroscopy; Models, Molecular; Molecular Sequence Data; Oxidation-Reduction; Protein Conformation; Protein Structure, Secondary; Solutions

The structure of an electron transfer complex containing a cytochrome c and a peroxidase - Pettigrew, G. W.; Prazeres, S.; Costa, C.; Palma, N.; Krippahl, L.; Moura, I.; Moura, J. J. (J Biol Chem, 1999)
Efficient biological electron transfer may require a fluid association of redox partners. Two noncrystallographic methods (a new molecular docking program and 1H NMR spectroscopy) have been used to study the electron transfer complex formed between the cytochrome c peroxidase (CCP) of Paracoccus denitrificans and cytochromes c. For the natural redox partner, cytochrome c550, the results are consistent with a complex in which the heme of a single cytochrome lies above the exposed electron-transferring heme of the peroxidase. In contrast, two molecules of the nonphysiological but kinetically competent horse cytochrome bind between the two hemes of the peroxidase. These dramatically different patterns are consistent with a redox active surface on the peroxidase that may accommodate more than one cytochrome and allow lateral mobility.
0021-9258 (Print)0021-9258 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Animals; Cytochrome c Group/*chemistry/metabolism; Electron Transport; Horses; Magnetic Resonance Spectroscopy; Peroxidases/*chemistry/metabolism; Protein Binding; Protein Conformation; Protons

Crystallization and preliminary x-ray analysis of a nitrate reductase from Desulfovibrio desulfuricans ATCC 27774 - Dias, J. M.; Bursakov, S.; Carneiro, C.; Moura, J. J.; Moura, I.; Romao, M. J. (Acta Crystallogr D Biol Crystallogr, 1999)
Periplasmic nitrate reductase from the sulfate-reducing bacterium Desulfovibrio desulfuricans ATCC 27774 contains two molybdopterin guanine dinucleotide cofactors and one [4Fe-4S] cluster as prosthetic groups and catalyzes the conversion of nitrate to nitrite. Crystals of the oxidized form of this enzyme were obtained using PEG as precipitant and belong to space group P3121 or P3221, with unit-cell dimensions a = b = 106.3, c = 135.1 A. There is one monomer of 80 kDa in the asymmetric unit, which corresponds to a Matthews ratio of 2.75 A3 Da-1. Using cryo-cooling procedures and X-rays from a rotating-anode generator, diffraction was observed to beyond 3.0 A resolution.
0907-4449 (Print)0907-4449 (Linking)Journal ArticleResearch Support, Non-U.S. Gov't
Keywords: Bacterial Proteins/*chemistry/isolation & purification; Crystallization; Crystallography, X-Ray; Desulfovibrio/*enzymology; Nitrate Reductase; Nitrate Reductases/*chemistry/isolation & purification

Enzymatic spectrophotometric determination of nitrites in beer - Girotti, S.; Ferri, E. N.; Fini, F.; Ruffini, F.; Budini, R.; Moura, I.; Almeida, G.; Costa, C.; Moura, J. J. G.; Carrea, G. (Analytical Letters, 1999)
A colorimetric assay for nitrite determination in beer based on c-type multiheme enzyme Nitrite reductase (NiR) isolated from Desulfovibrio desulfuricans ATCC 27774, was developed. Using the enzyme in solution, nitrite assay was linear in the 10(-8) - 10(-2) M range with a detection limit of 10(-8) M. and a recovery ranging from 90 to 107%. The imprecision ranged from 4 to 10% on the entire calibration curve. With NIR immobilised onto a nylon coil, a flow reactor was developed which showed a narrower linear range (10(-5) - 10(-2) M) and a higher detection limit (10(-5) M) than with the enzyme in solution, but made it possible to reuse the enzyme up to 100 times (50% residual activity). Sample preparation was simple and fast: only degassing and beer dilution by buffer was needed. This enzymatic assay was in good agreement with the results obtained using commercial nitrite determination kits.
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