Crystal Structures of a Family 19 Chitinase from Brassica Juncea Show Flexibility of Binding Cleft Loops.
From: Department of Molecular Biology, Swedish University of Agricultural Sciences, Uppsala, Sweden.
The FEBS journal
- Publish Date: Jul 2007
- ISSN: 1742-464X
- Volume: 274
- Issue: 14
- Pages: 3695-703
- Medium: Print
- Language: English
- Citation (JAMA): Ubhayasekera Wimal, Tang Ce Mun, Ho Sharon W T, et al. Crystal Structures of a Family 19 Chitinase from Brassica Juncea Show Flexibility of Binding Cleft Loops.. FEBS J. Jul 2007;274:3695-703
Abstract
Brassica juncea chitinase is an endo-acting, pathogenesis-related protein that is classified into glycoside hydrolase family 19, with highest homology (50-60%) in its catalytic domain to class I plant chitinases. Here we report X-ray structures of the chitinase catalytic domain from wild-type (apo, as well as with chloride ions bound) and a Glu234Ala mutant enzyme, solved by molecular replacement and refined at 1.53, 1.8 and 1.7 A resolution, respectively. Confirming our earlier mutagenesis studies, the active-site residues are identified as Glu212 and Glu234. Glu212 is believed to be the catalytic acid in the reaction, whereas Glu234 is thought to have a dual role, both activating a water molecule in its attack on the anomeric carbon, and stabilizing the charged intermediate. The molecules in the various structures differ significantly in the conformation of a number of loops that border the active-site cleft. The differences suggest an opening and closing of the enzyme during the catalytic cycle. Chitin is expected to dock first near Glu212, which will protonate it. Conformational changes then bring Glu234 closer, allowing it to assist in the following steps. These observations provide important insights into catalysis in family 19 chitinases.
Mesh Headings (Keywords): Binding Sites, Brassica, Chitinase, Crystallography, X-Ray, Models, Molecular, Protein Structure, Tertiary, Structural Homology, Protein
Check for Full Text / PubMed Unique Identifier (PMID): 17608716
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