Acta Crystallographica Section D

Biological Crystallography

Volume 69, Part 10 (October 2013)

research papers

Acta Cryst. (2013). D69, 2027-2038    [ doi:10.1107/S090744491301799X ]

The structure of a glycoside hydrolase family 81 endo-[beta]-1,3-glucanase

P. Zhou, Z. Chen, Q. Yan, S. Yang, R. Hilgenfeld and Z. Jiang

Abstract: Endo-[beta]-1,3-glucanases catalyze the hydrolysis of [beta]-1,3-glycosidic linkages in glucans. They are also responsible for rather diverse physiological functions such as carbon utilization, cell-wall organization and pathogen defence. Glycoside hydrolase (GH) family 81 mainly consists of [beta]-1,3-glucanases from fungi, higher plants and bacteria. A novel GH family 81 [beta]-1,3-glucanase gene (RmLam81A) from Rhizomucor miehei was expressed in Escherichia coli. Purified RmLam81A was crystallized and the structure was determined in two crystal forms (form I-free and form II-Se) at 2.3 and 2.0 Å resolution, respectively. Here, the crystal structure of a member of GH family 81 is reported for the first time. The structure of RmLam81A is greatly different from all endo-[beta]-1,3-glucanase structures available in the Protein Data Bank. The overall structure of the RmLam81A monomer consists of an N-terminal [beta]-sandwich domain, a C-terminal ([alpha]/[alpha])6 domain and an additional domain between them. Glu553 and Glu557 are proposed to serve as the proton donor and basic catalyst, respectively, in a single-displacement mechanism. In addition, Tyr386, Tyr482 and Ser554 possibly contribute to both the position or the ionization state of the basic catalyst Glu557. The first crystal structure of a GH family 81 member will be helpful in the study of the GH family 81 proteins and endo-[beta]-1,3-glucanases.

PDB references: 4k3a and 4k35

Keywords: glycoside hydrolases; family 81; endo-[beta]-1,3-glucanases; Rhizomucor miehei; three-dimensional structure.

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