Structural basis of lipopolysaccharide maturation by the O-antigen ligase.

Autor: Ashraf KU; Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, NY, USA., Nygaard R; Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, NY, USA., Vickery ON; School of Life Sciences, University of Warwick, Coventry, UK.; Department of Chemistry, University of Warwick, Coventry, UK., Erramilli SK; Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL, USA., Herrera CM; Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA, USA., McConville TH; Department of Medicine, Division of Infectious Diseases, Columbia University Medical Center, New York, NY, USA., Petrou VI; Department of Microbiology, Biochemistry, and Molecular Genetics, New Jersey Medical School, Rutgers Biomedical Health Sciences, Newark, NJ, USA.; Center for Immunity and Inflammation, New Jersey Medical School, Rutgers Biomedical Health Sciences, Newark, NJ, USA., Giacometti SI; Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, NY, USA., Dufrisne MB; Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, NY, USA., Nosol K; Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL, USA., Zinkle AP; Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, NY, USA., Graham CLB; School of Life Sciences, University of Warwick, Coventry, UK., Loukeris M; New York Consortium on Membrane Protein Structure, New York Structural Biology Center, New York, NY, USA., Kloss B; New York Consortium on Membrane Protein Structure, New York Structural Biology Center, New York, NY, USA., Skorupinska-Tudek K; Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland., Swiezewska E; Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland., Roper DI; Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, NY, USA.; School of Life Sciences, University of Warwick, Coventry, UK., Clarke OB; Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, NY, USA.; Department of Anesthesiology, Columbia University Irving Medical Center, New York, NY, USA., Uhlemann AC; Department of Medicine, Division of Infectious Diseases, Columbia University Medical Center, New York, NY, USA., Kossiakoff AA; Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL, USA., Trent MS; Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA, USA. strent@uga.edu., Stansfeld PJ; School of Life Sciences, University of Warwick, Coventry, UK. phillip.stansfeld@warwick.ac.uk.; Department of Chemistry, University of Warwick, Coventry, UK. phillip.stansfeld@warwick.ac.uk., Mancia F; Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center, New York, NY, USA. fm123@cumc.columbia.edu.
Jazyk: angličtina
Zdroj: Nature [Nature] 2022 Apr; Vol. 604 (7905), pp. 371-376. Date of Electronic Publication: 2022 Apr 06.
DOI: 10.1038/s41586-022-04555-x
Abstrakt: The outer membrane of Gram-negative bacteria has an external leaflet that is largely composed of lipopolysaccharide, which provides a selective permeation barrier, particularly against antimicrobials 1 . The final and crucial step in the biosynthesis of lipopolysaccharide is the addition of a species-dependent O-antigen to the lipid A core oligosaccharide, which is catalysed by the O-antigen ligase WaaL 2 . Here we present structures of WaaL from Cupriavidus metallidurans, both in the apo state and in complex with its lipid carrier undecaprenyl pyrophosphate, determined by single-particle cryo-electron microscopy. The structures reveal that WaaL comprises 12 transmembrane helices and a predominantly α-helical periplasmic region, which we show contains many of the conserved residues that are required for catalysis. We observe a conserved fold within the GT-C family of glycosyltransferases and hypothesize that they have a common mechanism for shuttling the undecaprenyl-based carrier to and from the active site. The structures, combined with genetic, biochemical, bioinformatics and molecular dynamics simulation experiments, offer molecular details on how the ligands come in apposition, and allows us to propose a mechanistic model for catalysis. Together, our work provides a structural basis for lipopolysaccharide maturation in a member of the GT-C superfamily of glycosyltransferases.
(© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)
Databáze: MEDLINE
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