Acceptor Substrate Discrimination in Phosphatidyl-myo-inositol Mannoside Synthesis STRUCTURAL AND MUTATIONAL ANALYSIS OF MANNOSYLTRANSFERASE CORYNEBACTERIUM GLUTAMICUM PimB′

被引：31

作者：

Batt, Sarah M. ^{[1
]}

Jabeen, Talat ^{[1
]}

Mishra, Arun K. ^{[1
]}

Veerapen, Natacha ^{[1
]}

Krumbach, Karin ^{[2
]}

Eggeling, Lothar ^{[2
]}

Besra, Gurdyal S. ^{[1
]}

Fuetterer, Klaus ^{[1
]}

机构：

[1] Univ Birmingham, Sch Biosci, Birmingham B15 2TT, W Midlands, England

[2] Forschungszentrum Julich, Inst Biotechnol 1, D-52425 Julich, Germany

来源：

JOURNAL OF BIOLOGICAL CHEMISTRY | 2010年 / 285卷 / 48期

基金：

英国惠康基金;

关键词：

MYCOBACTERIUM-TUBERCULOSIS; BIOSYNTHESIS; GLYCOSYLTRANSFERASES; PIMA; IDENTIFICATION; REFINEMENT; CATALYSIS; INSIGHTS; SYNTHASE; REVEALS;

D O I：

10.1074/jbc.M110.165407

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：

Long term survival of the pathogen Mycobacterium tuberculosis in humans is linked to the immunomodulatory potential of its complex cell wall glycolipids, which include the phosphatidylinositol mannoside (PIM) series as well as the related lipomannan and lipoarabinomannan glycoconjugates. PIM biosynthesis is initiated by a set of cytosolic alpha-mannosyltransferases, catalyzing glycosyl transfer from the activated saccharide donor GDP-alpha-D-mannopyranose to the acceptor phosphatidyl-myoinositol (PI) in an ordered and regio-specific fashion. Herein, we report the crystal structure of mannosyltransferase Corynebacterium glutamicum PimB' in complex with nucleotide to a resolution of 2.0 A. PimB' attaches mannosyl selectively to the 6-OH of the inositol moiety of PI. Two crystal forms and GD-Pversus GDP-alpha-D-mannopyranose-bound complexes reveal flexibility of the nucleotide conformation as well as of the structural framework of the active site. Structural comparison, docking of the saccharide acceptor, and site-directed mutagenesis pin regio-selectivity to a conserved Asp residue in the N-terminal domain that forces presentation of the correct inositol hydroxyl to the saccharide donor.

引用

页码：37741 / 37752

页数：12

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