Substrate-guided front-face reaction revealed by combined structural snapshots and metadynamics for the polypeptide N-acetylgalactosaminyltransferase 2
- Lira-Navarrete, E. 1
- Iglesias-Fernández, J. 3
- Zandberg, W.F. 5
- Compañón, I. 4
- Kong, Y. 2
- Corzana, F. 4
- Pinto, B.M. 5
- Clausen, H. 2
- Peregrina, J.M. 4
- Vocadlo, D.J. 5
- Rovira, C. 36
- Hurtado-Guerrero, R. 1
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1
Universidad de Zaragoza
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2
University of Copenhagen
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3
Universitat de Barcelona
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4
Universidad de La Rioja
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5
Simon Fraser University
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6
Institució Catalana de Recerca i Estudis Avançats
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ISSN: 1433-7851
Argitalpen urtea: 2014
Alea: 53
Zenbakia: 31
Orrialdeak: 8206-8210
Mota: Artikulua
Beste argitalpen batzuk: Angewandte Chemie International
Lotura duten proiektuak
Laburpena
The retaining glycosyltransferase GalNAc-T2 is a member of a large family of human polypeptide GalNAc-transferases that is responsible for the post-translational modification of many cell-surface proteins. By the use of combined structural and computational approaches, we provide the first set of structural snapshots of the enzyme during the catalytic cycle and combine these with quantum-mechanics/molecular-mechanics (QM/MM) metadynamics to unravel the catalytic mechanism of this retaining enzyme at the atomic-electronic level of detail. Our study provides a detailed structural rationale for an ordered bi-bi kinetic mechanism and reveals critical aspects of substrate recognition, which dictate the specificity for acceptor Thr versus Ser residues and enforce a front-face SNi-type reaction in which the substrate N-acetyl sugar substituent coordinates efficient glycosyl transfer. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.