3w7x

Crystal structure of E. coli YgjK D324N complexed with melibiose

spinqualitylabelsall models 3w7x, resolution 2.70Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 3w7x is a 2 chain structure with sequence from Escherichia coli K-12. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 2.7Å Ligands: CA, GLA, GLC, PRD_900037 Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function YGJK_ECOLI Glucoside hydrolase that cleaves the alpha-1,3-glucosidic linkage in nigerose. Has very low activity towards maltooligosaccharides, soluble starch, nigerotriose, kojibiose and trehalose. Publication Abstract from PubMed Proteins belonging to the glycoside hydrolase family 63 (GH63) are found in bacteria, archaea, and eukaryotes. Although the eukaryotic GH63 proteins have been identified as processing alpha-glucosidase I, the substrate specificities of the bacterial and archaeal GH63 proteins are not clear. Here, we converted a bacterial GH63 enzyme, Escherichia coli YgjK, to a glycosynthase to probe its substrate specificity. Two mutants of YgjK (E727A and D324N) were constructed, and both mutants showed glycosynthase activity. The reactions of E727A with beta-D-glucosyl fluoride and monosaccharides showed that the largest amount of glycosynthase product accumulated when galactose was employed as an acceptor molecule. The crystal structure of E727A complexed with the reaction product indicated that the disaccharide bound at the active site was 2-O-alpha-D-glucopyranosyl-alpha-D-galactopyranose (Glc12Gal). A comparison of the structures of E727A-Glc12Gal and D324N-melibiose showed that there were largely two types of conformations, which were the open and closed forms. The structure of YgjK adopted the closed form when the subsite -1 was occupied by glucose. These results suggest that sugars containing the Glc12Gal structure are the most likely candidates for natural substrates of YgjK. This article is protected by copyright. All rights reserved. Structure of a bacterial glycoside hydrolase family 63 enzyme in complex with its glycosynthase product and insights into the substrate specificity.,Miyazaki T, Ichikawa M, Yokoi G, Kitaoka M, Mori H, Kitano Y, Nishikawa A, Tonozuka T FEBS J. 2013 Jul 4. doi: 10.1111/febs.12424. PMID:23826932 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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