Structural Insights into the Substrate Specificity of a 6-Phospho-β-glucosidase BglA-2 from Streptococcus pneumoniae TIGR4
Year: 2013
Type: article
Abstract: The 6-phospho-β-glucosidase BglA-2 (EC 3.2.1.86) from glycoside hydrolase family 1 (GH-1) catalyzes the hydrolysis of β-1,4-linked cellobiose 6-phosphate (cellobiose-6′P) to yield glucose and glucose 6-phosphate. Both reaction products are further metabolized by the energy-generating glycolytic pathway. Here, we present the first crystal structures of the apo and complex forms of BglA-2 with thiocellobiose-6′P (a non-metabolizable analog of cellobiose-6′P) at 2.0 and 2.4 Å resolution, respectively. Similar to other GH-1 enzymes, the overall structure of BglA-2 from Streptococcus pneumoniae adopts a typical (β/α) 8 TIM-barrel, with the active site located at the center of the convex surface of the β-barrel. Structural analyses, in combination with enzymatic data obtained from site-directed mutant proteins, suggest that three aromatic residues, Tyr 126 , Tyr 303 , and Trp 338 , at subsite +1 of BglA-2 determine substrate specificity with respect to 1,4-linked 6-phospho-β-glucosides. Moreover, three additional residues, Ser 424 , Lys 430 , and Tyr 432 of BglA-2, were found to play important roles in the hydrolytic selectivity toward phosphorylated rather than non-phosphorylated compounds. Comparative structural analysis suggests that a tryptophan versus a methionine/alanine residue at subsite −1 may contribute to the catalytic and substrate selectivity with respect to structurally similar 6-phospho-β-galactosidases and 6-phospho-β-glucosidases assigned to the GH-1 family. Background: Streptococcus pneumoniae BglA-2 is a GH-1 6-phospho-β-glucosidase with specificity toward 1,4-linked 6-phospho-β-glucosides. Results: BglA-2 and other GH-1 members adopt a similar overall structure and catalytic mechanism. Conclusion: Tyr 126 , Tyr 303 , and Trp 338 determine substrate specificity, and Ser 424 , Lys 430 , and Tyr 432 discriminate phosphorylated from non-phosphorylated substrate. A tryptophan residue discriminates 6-phospho-β-glucosidase from 6-phospho-β-galactosidase activities. Significance: BglA-2 structures provide new insight into characteristics and substrate specificity of 6-phospho-β-glucosidase.
Source: Journal of Biological Chemistry
Authors: Wei-Li Yu, Yong-Liang Jiang, Andreas Pikis, Wang Cheng, Xiao-Hui Bai, Yan-Min Ren, John Thompson, Cong-Zhao Zhou, Yuxing Chen
Language: English
Subfield: Materials Chemistry
Field: Materials Science
Domain: Physical Sciences
Open Access status: hybrid
Estimated APC paid: $3,430
OpenAlex ID: W2016509969