A Native Ternary Complex Trapped in a Crystal Reveals the Catalytic Mechanism of a Retaining Glycosyltransferase

被引：35

作者：

Albesa-Jove, David ^{[1
,2
,10
]}

Mendoza, Fernanda ^{[4
,5
]}

Rodrigo-Unzueta, Ane ^{[1
,2
]}

Gomollon-Bel, Fernando ^{[8
]}

Cifuente, Javier O. ^{[1
,2
]}

Urresti, Saioa ^{[1
,2
]}

Comino, Natalia ^{[1
,2
]}

Gomez, Hansel ^{[6
,7
]}

Romero-Garcia, Javier ^{[9
]}

Lluch, Jose M. ^{[4
,5
]}

Sancho-Vaello, Enea ^{[1
,2
]}

Biarnes, Xevi ^{[9
]}

Planas, Antoni ^{[9
]}

Merino, Pedro ^{[8
]}

Masgrau, Laura ^{[3
]}

Guerin, Marcelo E. ^{[1
,2
,10
]}

机构：

[1] Univ Basque Country, Unidad Biofis, Consejo Super Invest Cient, Euskal Herriko Unibertsitatea CSIC UPV EHU, Leioa 48940, Bizkaia, Spain

[2] Univ Basque Country, Dept Bioquim, Leioa 48940, Bizkaia, Spain

[3] IBB, Minorca, Spain

[4] Univ Autonoma Barcelona, IBB, Bellaterra 08193, Spain

[5] Univ Autonoma Barcelona, Dept Quim, Bellaterra 08193, Spain

[6] IRB Barcelona, IBB, Barcelona 08028, Spain

[7] IRB Barcelona, Joint BSC CRG IRB Program Computat Biol, Barcelona 08028, Spain

[8] Univ Zaragoza, Lab Sintesis Asimtr, Dept Sintesis & Estruct Biomol, ISQCH, Zaragoza 50009, Aragon, Spain

[9] Univ Ramon Llull, Biochem Lab, Inst Quim Sarria, Barcelona 08017, Spain

[10] Ikerbasque, Bilbao 48013, Spain

来源：

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION | 2015年 / 54卷 / 34期

关键词：

enzyme catalysis; enzymes; glycosyltransferases; reaction mechanisms; structure elucidation; SUBSTRATE; INSIGHTS; MUTANT; DONOR;

D O I：

10.1002/anie.201504617

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Glycosyltransferases (GTs) comprise a prominent family of enzymes that play critical roles in a variety of cellular processes, including cell signaling, cell development, and host-pathogen interactions. Glycosyl transfer can proceed with either inversion or retention of the anomeric configuration with respect to the reaction substrates and products. The elucidation of the catalytic mechanism of retaining GTs remains a major challenge. A native ternary complex of a GT in a productive mode for catalysis is reported, that of the retaining glucosyl-3-phosphoglycerate synthase GpgS from M. tuberculosis in the presence of the sugar donor UDP-Glc, the acceptor substrate phosphoglycerate, and the divalent cation cofactor. Through a combination of structural, chemical, enzymatic, molecular dynamics, and quantum-mechanics/molecular-mechanics (QM/MM) calculations, the catalytic mechanism was unraveled, thereby providing a strong experimental support for a front-side substrate-assisted SNi-type reaction.

引用

页码：9898 / 9902

页数：5

共 25 条

[1] Structure-function relationships of membrane-associated GT-B glycosyltransferases [J].