Mass spectrometric characterization of a two-glycosyltransferase tandem reaction for assembly of tetrasaccharide repeating unit of Escherichia coli O77 O-antigen

被引：3
作者：
Zhou, Dawei [1 ]
Chen, Chao [1 ]
Xu, Lingling [1 ]
Utkina, Natalia [2 ]
Danilov, Leonid [2 ]
Torgov, Vladimir [2 ]
Veselovsky, Vladimir [2 ]
Liu, Bin [1 ]
Feng, Lu [1 ]
机构：
[1] Nankai Univ, TEDA Inst Biol Sci & Biotechnol, TEDA, 23 Hongda St, Tianjin 300457, Peoples R China
[2] Russian Acad Sci, ND Zelinsky Inst Organ Chem, Leninsky Prosp 47, Moscow 119991, Russia
来源：
CARBOHYDRATE RESEARCH | 2016年 / 424卷
基金：
中国国家自然科学基金; 对外科技合作项目（国际科技项目）;
关键词：
E. coli O77; O-antigen repeating unit; Glycosyltransferases; WbaD; WbaC; Electrospray ionization multistage tandem mass spectrometry; BIOCHEMICAL-CHARACTERIZATION; BIOSYNTHESIS; IDENTIFICATION; OLIGOSACCHARIDES; ARABINOFURANOSYLTRANSFERASE; LIPOPOLYSACCHARIDE; POLYSACCHARIDE; ASSOCIATIONS; SEROGROUPS; TOXINS;
D O I：
10.1016/j.carres.2016.02.007
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：
The wbaD gene and wbaC gene from Escherichia coli O77 O-antigen gene cluster encoding mannosyltransferases were functionally characterized in vitro. A synthetic acceptor P-1-(11-phenoxyundecyl)-P-2-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl) diphosphate (GlcNAc-PP-PhU) was used as an acceptor and GDP-Man as a donor substrate; the activities of WbaD and WbaC were confirmed by detailed structural characterization of their lipooligosacharide enzyme products using high-sensitivity negative-ion electrospray ionization (ESI) collision-induced dissociation tandem mass spectrometry (CID) MS-MS. The extensive fragmentation unequivocally demonstrated that the Man(1-3)-GlcNAc linkage in WbaD catalyzed reaction product and two Man(1-2)-Man linkages in tandem WbaD/WbaC catalyzed reaction product are present, respectively. This study provided valuable information for the understanding of diversified glycosyltransferase (GT) functions and the two GTs characterized can serve as additional enzyme sources for possible pharmaceutical related applications. (C) 2016 Elsevier Ltd. All rights reserved.
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页码：24 / 29
页数：6
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