DNA damage and metal accumulation in four tissues of feral Octopus vulgaris from two coastal areas in Portugal,
Raimundo, Joana, Costa Pedro M., Vale Carlos, Costa Maria Helena, and Moura Isabel
, Ecotoxicology and Environmental Safety, Oct, Volume 73, Number 7, p.1543-1547, (2010)
AbstractThe alkaline comet assay has been employed for the first time to estimate the basal DNA damage in the digestive gland, gills, kidney and gonads of Octopus vulgaris. Octopuses were captured in two coastal areas adjacent to the cities of Matosinhos (N) and Olhao (S), Portugal. The area of Matosinhos is influenced by discharges of the Douro River, city of Porto, industries and intensive agriculture, while Olhao is an important fisheries port. Previous works point to contrasting metal availability in the two coastal areas. Among the analysed tissues digestive gland presented the highest levels of Zn, Cu, Cd and Pb. Tissues of specimens from Matosinhos exhibited high levels of Cd and from Olhao enhanced Pb concentrations. The DNA damages in digestive gland, gills and kidney were more accentuated in specimens from Matosinhos than from Olhao, suggesting a stronger effect of contaminants. Elevated strand breakages were registered in digestive gland, recognised for its ability to store and detoxify accumulated metals. The DNA damages in kidney, gills and gonads were lower, reflecting reduced metal accumulation or efficient detoxification. The broad variability of damages in the three tissues may also mirror tissue function, specific defences to genotoxicants and cell-cycle turnover. (C) 2010 Elsevier Inc. All rights reserved.
Association of Zn, Cu, Cd and Pb with protein fractions and sub-cellular partitioning in the digestive gland of Octopus vulgaris living in habitats with different metal levels,
Raimundo, J., Vale C., Duarte R., and Moura I.
, Chemosphere, Nov, Volume 81, Number 10, p.1314-1319, (2010)
AbstractZinc Cu Cd and Pb concentrations were determined in protein fractions of digestive gland and in the whole digestive gland of Octopus vulgaris collected from two areas of the Portuguese coast Approximately 95% of Zn 99% of Cu 85-96% of Cd and 77-86% of Pb were stored in the cytosol suggesting the predominance of cytosolic proteins in the trapping these elements Gel filtration chromatography evidenced the presence of two major groups of proteins with high molecular weight (HMW 144 000-130 000 Da) and low molecular weight (LMW 11 000-6000 Da) The following metal-protein associations were found Zn was distributed between HMW and LMW Cu and Cd in LMW proteins with a minor association with HMW and Pb in HMW proteins The strong positive correlations between Cd Zn and Cu and LMW proteins point to the presence of metalloproteins with high affinity to these elements A shift was registered between the maximum of the ratio 254 280 nm and metal concentrations in the chromatographic profiles This shift may result from metallothioneins having a small participation in the metal binding or protein purification was insufficient and various LMW proteins may be interfering (C) 2010 Elsevier Ltd All rights reserved
The 1.4 angstrom resolution structure of Paracoccus pantotrophus pseudoazurin,
Najmudin, Shabir, Pauleta Sofia R., Moura Isabel, and Romao Maria J.
, Acta Crystallographica Section F-Structural Biology and Crystallization Communications, Jun, Volume 66, p.627-635, (2010)
AbstractPseudoazurins are small type 1 copper proteins that are involved in the flow of electrons between various electron donors and acceptors in the bacterial periplasm, mostly under denitrifying conditions. The previously determined structure of Paracoccus pantotrophus pseudoazurin in the oxidized form was improved to a nominal resolution of 1.4 angstrom, with R and R(free) values of 0.188 and 0.206, respectively. This high-resolution structure makes it possible to analyze the interactions between the monomers and the solvent structure in detail. Analysis of the high-resolution structure revealed the structural regions that are responsible for monomer-monomer recognition during dimer formation and for protein-protein interaction and that are important for partner recognition. The pseudoazurin structure was compared with other structures of various type 1 copper proteins and these were grouped into families according to similarities in their secondary structure; this may be useful in the annotation of copper proteins in newly sequenced genomes and in the identification of novel copper proteins.
Implications of oxidovanadium(IV) binding to actin,
Ramos, S., Almeida R. M., Moura J. J., and Aureliano M.
, J Inorg Biochem, Jun, Volume 105, Number 6, p.777-83, (2010)
AbstractOxidovanadium(IV), a cationic species (VO(2+)) of vanadium(IV), binds to several proteins, including actin. Upon titration with oxidovanadium(IV), approximately 100% quenching of the intrinsic fluorescence of monomeric actin purified from rabbit skeletal muscle (G-actin) was observed, with a V(50) of 131 muM, whereas for the polymerized form of actin (F-actin) 75% of quenching was obtained and a V(50) value of 320 muM. Stern-Volmer plots were used to estimate an oxidovanadium(IV)-actin dissociation constant, with K(d) of 8.2 muM and 64.1 muM VOSO(4), for G-actin and F-actin, respectively. These studies reveal the presence of a high affinity binding site for oxidovanadium(IV) in actin, producing local conformational changes near the tryptophans most accessible to water in the three-dimensional structure of actin. The actin conformational changes, also confirmed by (1)H NMR, are accompanied by changes in G-actin hydrophobic surface, but not in F-actin. The (1)H NMR spectra of G-actin treated with oxidovanadium(IV) clearly indicates changes in the resonances ascribed to methyl group and aliphatic regions as well as to aromatics and peptide-bond amide region. In parallel, it was verified that oxidovanadium(IV) prevents the G-actin polymerization into F-actin. In the 0-200 muM range, VOSO(4) inhibits 40% of the extent of polymerization with an IC(50) of 15.1 muM, whereas 500 muM VOSO(4) totally suppresses actin polymerization. The data strongly suggest that oxidovanadium(IV) binds to actin at specific binding sites preventing actin polymerization. By affecting actin structure and function, oxidovanadium(IV) might be responsible for many cellular effects described for vanadium.
Relations between mercury, methyl-mercury and selenium in tissues of Octopus vulgaris from the Portuguese Coast,
Raimundo, Joana, Vale Carlos, Canario Joao, Branco Vasco, and Moura Isabel
, Environmental Pollution, Jun, Volume 158, Number 6, p.2094-2100, (2010)
AbstractMercury, methyl-mercury (MeHg) and selenium were determined in digestive gland and mantle of Octopus vulgaris, from three areas of the Portuguese coast. To our knowledge these are the first data on MeHg in cephalopods. Concentrations were higher in the digestive gland and percentage of MeHg in mantle. Enhanced Hg and MeHg levels were obtained in digestive gland of specimens from Olhao (3.1-7.4 and 2.0-5.0 mu g g(-1) respectively). Differences between areas may be partially related to Hg availability. Relationships between concentrations in mantle and digestive gland pointed to proportional increases of Hg and MeHg in tissues of specimens from Matosinhos and Cascais, but relatively constant values in mantle of individuals from Olhao (higher contamination). Se:Hg molar ratio in digestive gland was 32 and 30 in octopus from Matosinhos and Cascais, respectively, and 5.4 from Olhao. The proximity to the unit suggests demethylation as response to elevated MeHg levels in digestive gland. (C) 2010 Elsevier Ltd. All rights reserved.
Metallothioneins and trace elements in digestive gland, gills, kidney and gonads of Octopus vulgaris,
Raimundo, J., Costa P. M., Vale C., Costa M. H., and Moura I.
, Comparative Biochemistry and Physiology C-Toxicology & Pharmacology, Aug, Volume 152, Number 2, p.139-146, (2010)
AbstractMetallothionein-like proteins (MT) and V, Cr, Co, Ni, Zn, Cu, As and Cd were determined in digestive gland, gills, kidney and gonads of Octopus vulgaris, from the Portuguese coast. To our knowledge these are the first data on MT in octopus. High concentrations (mu g g(-1), dry mass) of Zn (48050) and Cd (555) were found in digestive gland, and MT reached levels one order of magnitude above the ones registered in wild bivalves. Significantly higher levels of MT in digestive gland and gills of specimens from A and B were in line with elevated Cd concentrations. Principal component analyses (PCA) point to MT-Cd and MT-Cr associations in digestive gland and gills. Despite the high levels of Zn in specimens from B, association with Zn was not obtained. Due to the affinity of MT to various elements, it should not be excluded the possibility of Cd replacing Zn in Zn-MT. Kidney presented higher levels of Cd, Co, Ni and As than gills and gonads, and in the case of As surpassing the levels in digestive gland, but PCA showed no relation with MT. Likewise the MT levels in gonads had no correspondence to the metal concentration variation. (C) 2010 Elsevier Inc. All rights reserved.
Ultrasonic multiprobe as a new tool to overcome the bottleneck of throughput in workflows for protein identification relaying on ultrasonic energy,
Santos, H. M., Carreira R., Diniz M. S., Rivas M. G., Lodeiro C., Moura J. J., and Capelo J. L.
, Talanta, Apr 15, Volume 81, Number 1-2, p.55-62, (2010)
AbstractWe studied in this work the performance of the new ultrasonic multiprobe in terms of throughput, handling and robustness. The study was conducted using the multiprobe to speed two different proteomics workflows. The "classic" method relaying on overnight protein digestion (12h), was used as the standard procedure. This work clearly shows the importance of testing variables such as ultrasonic amplitude and ultrasonic time when adapting an ultrasonic-based treatment to a new ultrasonic device. The results here presented also shown and confirm the advantage of speed up sample treatment workflows with the aid of ultrasonic energy in combination with a 96-well plate. The methods compared were similar in terms of robustness, but the desalting free method was the fastest, requiring only 2 min/sample for completion. In addition it was also the simplest in terms of handling, since no desalting step was needed. The following standard proteins were successfully identified using the methods studied: bovine serum albumin, alpha-lactalbumin, ovalbumin, carbonic anhydrase, fructose-bisphosphate aldolase A, catalase, chymotrypsinogen A. As case study, the identification of the protein Split-Soret cytochrome c from D. desulfuricans ATCC 27774 was carried out.