@article {Pires2017a, title = {Stability and ligand promiscuity of type A carbohydrate-binding modules are illustrated by the structure of Spirochaeta thermophila StCBM64C}, journal = {Journal of Biological Chemistry}, volume = {292}, number = {12}, year = {2017}, note = {

n/a

}, month = {mar}, pages = {4847{\textendash}4860}, abstract = {

Deconstruction of cellulose, the most abundant plant cell wall polysaccharide, requires the cooperative activity of a large repertoire of microbial enzymes. Modular cellulases contain non-catalytic type A Carbohydrate-Binding Modules (CBMs) that specifically bind to the crystalline regions of cellulose, thus promoting enzyme efficacy through proximity and targeting effects. Although type A CBMs play a critical role in cellulose recycling, their mechanism of action remains poorly understood. Here we produced a library of recombinant CBMs representative of the known diversity of type A modules. The binding properties of 40 CBMs, in fusion with an N-terminal green fluorescence protein (GFP) domain, revealed that type A CBMs possess the ability to recognize different crystalline forms of cellulose and chitin over a wide range of temperatures, pHs and ionic strengths. A Spirochaeta thermophila CBM64, in particular, displayed plasticity in its capacity to bind both crystalline and soluble carbohydrates under a wide range of extreme conditions. The structure of S. thermophila StCBM64C revealed an untwisted, flat, carbohydrate-binding interface comprising the side chains of four tryptophan residues in a coplanar linear arrangement. Significantly, two highly conserved asparagine side chains, each one located between two tryptophan residues, are critical to insoluble and soluble glucan recognition but not to bind xyloglucan. Thus, CBM64 compact structure and its extended and versatile ligand interacting platform illustrates how type A CBMs target their appended plant cell wall degrading enzymes to a diversity of recalcitrant carbohydrates under a wide range of environmental conditions.

}, issn = {1083351X}, doi = {10.1074/jbc.M116.767541}, url = {http://europepmc.org/abstract/med/28179427}, author = {Pires, Virg{\'\i}nia M R and Pereira, Pedro M M and Br{\'a}s, Joana L A and Correia, M{\'a}rcia and Cardoso, V{\^a}nia and Bule, Pedro and Alves, Victor D. and Najmudin, Shabir and Venditto, Immacolata and Ferreira, Lu{\'\i}s M. A. and Rom{\~a}o, Maria Jo{\~a}o and Ana Lu{\'\i}sa Carvalho and Fontes, Carlos M. G. A. and Prazeres, Duarte Miguel} }