Mechanistic snapshots of lipid-linked sugar transfer

Enzymes undergo dynamic conformational changes during catalysis, yet conventional high-resolution structural methods typically capture only the most stable states. Here, we address this gap using rapid UV photolysis of a chemically caged substrate with cryogenic time-resolved electron microscopy (cryo-TREM). We elucidate the catalytic mechanism of GtrB, a membrane-bound glycosyltransferase that transfers glucose from UDP-glucose to the lipid carrier undecaprenyl phosphate. We visualized how GtrB, which has an active site ~15 Å from the membrane, transitions during the catalytic cycle to move each substrate in proximity for catalysis. From a single dataset, we resolved distinct conformational states: the initial substrate-bound state, a catalytically poised intermediate, and the product-bound state. Through molecular dynamics simulations and biochemical analyses, we identify coordinated movements within the active site that drive catalysis. These findings provide a molecular framework for understanding how glycosyltransferases function and highlight a broadly applicable strategy for capturing dynamic enzymatic states in native-like environments.

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Publication Details

Journal
Nature Communications
Published
2025-12-06
DOI
https://doi.org/10.1038/s41467-025-66769-7
Primary Topic
Lipid Membrane Structure and Behavior
Type
article
Field-Weighted Citation Impact
0.00

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article

Mechanistic snapshots of lipid-linked sugar transfer

Todd L. Lowary, Khuram U. Ashraf, Michael H. B. Stowell, Alice Romagnoli et al.
Nature Communications
Lipid Membrane Structure and Behavior
article

Mechanistic snapshots of lipid-linked sugar transfer

Todd L. Lowary, Khuram U. Ashraf, Michael H. B. Stowell, Alice Romagnoli, Matthias Quick, Filippo Mancia, Bedangshu Mishra, Ryan T. Morgan, Rie Nygaard, Biddut Bhattacharjee, Vasileios I. Petrou, Eva Gil‐Iturbe, Injin Bang, Giovanni Di Muccio, Stefano Motta, Manal M. Anwar, Daniele Di Marino
article en

Abstract

Enzymes undergo dynamic conformational changes during catalysis, yet conventional high-resolution structural methods typically capture only the most stable states. Here, we address this gap using rapid UV photolysis of a chemically caged substrate with cryogenic time-resolved electron microscopy (cryo-TREM). We elucidate the catalytic mechanism of GtrB, a membrane-bound glycosyltransferase that transfers glucose from UDP-glucose to the lipid carrier undecaprenyl phosphate. We visualized how GtrB, which has an active site ~15 Å from the membrane, transitions during the catalytic cycle to move each substrate in proximity for catalysis. From a single dataset, we resolved distinct conformational states: the initial substrate-bound state, a catalytically poised intermediate, and the product-bound state. Through molecular dynamics simulations and biochemical analyses, we identify coordinated movements within the active site that drive catalysis. These findings provide a molecular framework for understanding how glycosyltransferases function and highlight a broadly applicable strategy for capturing dynamic enzymatic states in native-like environments.

Nature CommunicationsVol. 16(1)
Rutgers, The State University of New Jersey (US), Marche Polytechnic University (IT), National Taiwan University (TW), University of Colorado Boulder (US), Mario Negri Institute for Pharmacological Research (IT), Cornell University (US), Columbia University Irving Medical Center (US), New York Psychoanalytic Society and Institute (US), Institute of Biological Chemistry, Academia Sinica (TW), Weill Cornell Medicine (US), Rutgers Health (US), University of Milano-Bicocca (IT)
European Commission, Academia Sinica, Università degli Studi di Milano-Bicocca, National Institute of General Medical Sciences
Openalex Percentile: Top 65%
Lipid Membrane Structure and Behavior
DOIOpen Access PDFOpenAlex
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