The κ-carrageenase of P-carrageenovora features a tunnel-shaped active site:: A novel insight in the evolution of clan-B glycoside hydrolases

被引:171
作者
Michel, G
Chantalat, L
Duee, E
Barbeyron, T
Henrissat, B
Kloareg, B
Dideberg, O
机构
[1] CEA, CNRS, Inst Biol Struct Jean Pierre Ebel, Lab Cristallog Macromol, F-38027 Grenoble, France
[2] CNRS, Biol Stn, UMR 1931, F-29682 Roscoff, France
[3] Lab Goemar, F-38027 Grenoble, France
[4] CNRS, IFR 1, F-13402 Marseille 20, France
关键词
glycoside hydrolase; MAD; sulfated galactan; X-ray structure;
D O I
10.1016/S0969-2126(01)00612-8
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Background: kappa -carrageenans are gel-forming, sulfated 1,3-alpha -1,4-beta -galactans from the cell walls of marine red algae. The kappa -carrageenase from the marine, gram-negative bacterium Pseudoalteromonas carrageenovora degrades kappa -carrageenan both in solution and in solid state by an endoprocessive mechanism. This beta -galactanase belongs to the clan-B of glycoside hydrolases. Results: The structure of P. carrageenovora kappa -carrageenase has been solved to 1.54 Angstrom resolution by the multiwavelength anomalous diffraction (MAD) method, using a seleno-methionine-substituted form of the enzyme. The enzyme folds into a curved beta sandwich, with a tunnel-like active site cavity. Another remarkable characteristic is the presence of an arginine residue at subsite -1. Conclusions: The crystal structure of P. carrageenovora kappa -carrageenase is the first three-dimensional structure of a carrageenase. Its tunnel-shaped active site, the first to be reported for enzymes other than cellulases, suggests that such tunnels are associated with the degradation of solid polysaccharides. Clan-B glycoside hydrolases fall into two subgroups, one with catalytic machinery held by an ancestral beta bulge, and the other in which it is held by a regular beta strand. At subsite -1,all of these hydrolases exhibit an aromatic amino acid that interacts with the hexopyranose ring of the monosaccharide undergoing catalysis. In addition, in kappa -carrageenases, an arginine residue recognizes the sulfate-ester substituents of the beta -linked kappa -carrageenan monomers. It also appears that, in addition to the nucleophile and acid/base catalysts, two other amino acids are involved with the catalytic cycle, accelerating the deglycosylation step.
引用
收藏
页码:513 / 525
页数:13
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