Structural studies on carbohydrate binding modules and ice-binding proteins

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Ab initio structure determination and functional characterization of CBM36: A new family of calcium-dependent carbohydrate binding molecules.

AbstractThe enzymatic degradation of polysaccharides harnesses multimodular enzymes whose carbohydrate binding modules (CBM) target the catalytic domain onto the recalcitrant substrate. Here we report the ab initio structure determination and subsequent refinement, at 0.8 Å resolution, of the CBM36 domain of the Paenibacillus polymyxa xylanase 43A. Affinity electrophoresis, isothermal titration calorimetry, and UV difference spectroscopy demonstrate that CBM36 is a novel Ca2+-dependent xylan binding domain. The 3D structure of CBM36 in complex with xylotriose and Ca2+, at 1.5 Å resolution, displays significant conformational changes compared to the native structure and reveals the molecular basis for its unique Ca2+-dependent binding of xylooligosaccharides through coordination of the O2 and O3 hydroxyls. CBM36 is one of an emerging spectrum of carbohydrate binding modules that increasingly find applications in industry and display great potential for mapping the “glyco-architecture” of plant cells