title: Identification of a broad family of lipid A late acyltransferases with non-canonical substrate specificity.
abstract: Most Gram-negative organisms produce lipopolysaccharide (LPS), a complex macromolecule anchored to the bacterial membrane by the lipid A moiety. Lipid A is synthesized via the Raetz pathway, a conserved nine-step enzymatic process first characterized in Escherichia coli. The Epsilonproteobacterium Helicobacter pylori uses the Raetz pathway to synthesize lipid A; however, only eight of nine enzymes in the pathway have been identified in this organism. Here, we identify the missing acyltransferase, Jhp0255, which transfers a secondary acyl chain to the 3'-linked primary acyl chain of lipid A, an activity similar to that of E. coli LpxM. This enzyme, reannotated as LpxJ due to limited sequence similarity with LpxM, catalyses addition of a C12:0 or C14:0 acyl chain to the 3'-linked primary acyl chain of lipid A, complementing an E. coli LpxM mutant. Enzymatic assays demonstrate that LpxJ and homologues in Campylobacter jejuni and Wolinella succinogenes can act before the 2' secondary acyltransferase, LpxL, as well as the 3-deoxy-d-manno-octulosonic acid (Kdo) transferase, KdtA. Ultimately, LpxJ is one member of a large class of acyltransferases found in a diverse range of organisms that lack an E. coli LpxM homologue, suggesting that LpxJ participates in lipid A biosynthesis in place of an LpxM homologue.
authors.affiliation: Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX, USA.
authors.forename: Erica J
authors.initials: EJ
authors.lastname: Rubin
authors.forename: John P
authors.initials: JP
authors.lastname: O'Brien
authors.forename: Petko L
authors.initials: PL
authors.lastname: Ivanov
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Brodbelt
authors.forename: M Stephen
authors.initials: MS
authors.lastname: Trent
mesh_terms: D000215:Acylation; D000217:Acyltransferases / Q000737:chemistry / Q000378:metabolism*; D001419:Bacteria / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D020565:Epsilonproteobacteria / Q000201:enzymology; D005816:Genetic Complementation Test; D008050:Lipid A / Q000737:chemistry / Q000378:metabolism*; D005810:Multigene Family; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D013400:Sugar Acids
chemical_list: D001426:Bacterial Proteins; D008050:Lipid A; D013400:Sugar Acids; C002532:2-keto-3-deoxyoctonate; D000217:Acyltransferases