Boraston Alisdair B, Nurizzo Didier, Notenboom Valerie, Ducros Valérie, Rose David R, Kilburn Douglas G, Davies Gideon J
Protein Engineering Network of Centres of Excellence, Edmonton, Alberta, Canada T6G 2S2.
J Mol Biol. 2002 Jun 21;319(5):1143-56. doi: 10.1016/S0022-2836(02)00374-1.
Enzymes active on complex carbohydrate polymers frequently have modular structures in which a catalytic domain is appended to one or more carbohydrate-binding modules (CBMs). Although CBMs have been classified into a number of families based upon sequence, many closely related CBMs are specific for different polysaccharides. In order to provide a structural rationale for the recognition of different polysaccharides by CBMs displaying a conserved fold, we have studied the thermodynamics of binding and three-dimensional structures of the related family 4 CBMs from Cellulomonas fimi Cel9B and Thermotoga maritima Lam16A in complex with their ligands, beta-1,4 and beta-1,3 linked gluco-oligosaccharides, respectively. These two CBMs use a structurally conserved constellation of aromatic and polar amino acid side-chains that interact with sugars in two of the five binding subsites. Differences in the length and conformation of loops in non-conserved regions create binding-site topographies that complement the known solution conformations of their respective ligands. Thermodynamics interpreted in the light of structural information highlights the differential role of water in the interaction of these CBMs with their respective oligosaccharide ligands.
作用于复合碳水化合物聚合物的酶通常具有模块化结构,其中催化结构域连接到一个或多个碳水化合物结合模块(CBM)上。尽管CBM已根据序列被分为多个家族,但许多密切相关的CBM对不同的多糖具有特异性。为了提供一个结构上的理论依据,以解释具有保守折叠的CBM对不同多糖的识别,我们研究了来自纤维单胞菌Cel9B和嗜热栖热菌Lam16A的相关4家族CBM与它们的配体(分别为β-1,4和β-1,3连接的葡萄糖寡糖)形成复合物时的结合热力学和三维结构。这两个CBM使用一组结构保守的芳香族和极性氨基酸侧链,它们在五个结合亚位点中的两个与糖相互作用。非保守区域中环的长度和构象的差异产生了与各自配体已知溶液构象互补的结合位点拓扑结构。根据结构信息解释的热力学突出了水在这些CBM与各自寡糖配体相互作用中的不同作用。