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

Publisher: Nature Publishing Group Country of Publication: England NLM ID: 0410462 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1476-4687 (Electronic) Linking ISSN: 00280836 NLM ISO Abbreviation: Nature Subsets: MEDLINE

Publication: Basingstoke : Nature Publishing Group
Original Publication: London, Macmillan Journals ltd.

Ligases* ; Lipopolysaccharides*/chemistry ; Lipopolysaccharides*/metabolism ; O Antigens*; Bacterial Proteins/chemistry ; Carbon-Oxygen Ligases/chemistry ; Carbon-Oxygen Ligases/genetics ; Cryoelectron Microscopy ; Glycosyltransferases ; Gram-Negative Bacteria

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.)

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0 (Bacterial Proteins)
0 (Lipopolysaccharides)
0 (O Antigens)
EC 2.4.- (Glycosyltransferases)
EC 6.- (Ligases)
EC 6.1.- (Carbon-Oxygen Ligases)

Date Created: 20220407 Date Completed: 20220415 Latest Revision: 20240822

20240823

PMC9884178

10.1038/s41586-022-04555-x

35388216