TY - JOUR
T1 - Structural Snapshots and Loop Dynamics along the Catalytic Cycle of Glycosyltransferase GpgS
AU - Albesa-Jové, David
AU - Romero-García, Javier
AU - Sancho-Vaello, Enea
AU - Contreras, F. Xabier
AU - Rodrigo-Unzueta, Ane
AU - Comino, Natalia
AU - Carreras-González, Ana
AU - Arrasate, Pedro
AU - Urresti, Saioa
AU - Biarnés, Xevi
AU - Planas, Antoni
AU - Guerin, Marcelo E.
N1 - Funding Information:
This work was supported by EU Contract HEALTH-F3-2011- 260872, MINECO Grant BIO2013-49022-C2-2-R, and the Basque Government (to M.E.G.), and MINECO Grant BIO2013-49022-C2-1-R (to A.P.) and BFU2012-33103 (to F.-X.C). J.R.G. acknowledges a predoctoral fellowship from IQS. We acknowledge Diamond Light Source (proposals mx8302/10130), SOLEIL (proposal 20140843), and BioStruct-X (proposals 2460/5594/7687) for providing access to synchrotron radiation facilities. Computational Resources were provided by the Red Española de Supercomputación (RES). We gratefully acknowledge S. López-Fernández (Unit of Biophysics, CSIC, UPV/EHU, Spain), and E. Ogando and T. Mercero (Scientific Computing Service UPV/EHU, Spain), for technical assistance. Technical and staff support provided by the Servicio de Lipidómica of the SGIker (UPV/EHU, MICINN, GV/EJ, ESF) is gratefully acknowledged. We also acknowledge all members of the Structural Glycobiology Group (Spain) for valuable scientific discussions.
Publisher Copyright:
© 2017 Elsevier Ltd
PY - 2017/7/5
Y1 - 2017/7/5
N2 - Glycosyltransferases (GTs) play a central role in nature. They catalyze the transfer of a sugar moiety to a broad range of acceptor substrates. GTs are highly selective enzymes, allowing the recognition of subtle structural differences in the sequences and stereochemistry of their sugar and acceptor substrates. We report here a series of structural snapshots of the reaction center of the retaining glucosyl-3-phosphoglycerate synthase (GpgS). During this sequence of events, we visualize how the enzyme guides the substrates into the reaction center where the glycosyl transfer reaction takes place, and unveil the mechanism of product release, involving multiple conformational changes not only in the substrates/products but also in the enzyme. The structural data are further complemented by metadynamics free-energy calculations, revealing how the equilibrium of loop conformations is modulated along these itineraries. The information reported here represent an important contribution for the understanding of GT enzymes at the molecular level.
AB - Glycosyltransferases (GTs) play a central role in nature. They catalyze the transfer of a sugar moiety to a broad range of acceptor substrates. GTs are highly selective enzymes, allowing the recognition of subtle structural differences in the sequences and stereochemistry of their sugar and acceptor substrates. We report here a series of structural snapshots of the reaction center of the retaining glucosyl-3-phosphoglycerate synthase (GpgS). During this sequence of events, we visualize how the enzyme guides the substrates into the reaction center where the glycosyl transfer reaction takes place, and unveil the mechanism of product release, involving multiple conformational changes not only in the substrates/products but also in the enzyme. The structural data are further complemented by metadynamics free-energy calculations, revealing how the equilibrium of loop conformations is modulated along these itineraries. The information reported here represent an important contribution for the understanding of GT enzymes at the molecular level.
KW - carbohydrate biosynthesis
KW - carbohydrate modifying enzymes
KW - enzyme catalysis
KW - enzyme dynamics
KW - enzyme mechanism
KW - enzyme structure
KW - glycosyltransferases
KW - lipopolysaccharide
KW - mycobacteria
UR - http://www.scopus.com/inward/record.url?scp=85020708114&partnerID=8YFLogxK
U2 - 10.1016/j.str.2017.05.009
DO - 10.1016/j.str.2017.05.009
M3 - Article
C2 - 28625787
AN - SCOPUS:85020708114
SN - 0969-2126
VL - 25
SP - 1034-1044.e3
JO - Structure
JF - Structure
IS - 7
ER -