title: Elucidation of a novel Vibrio cholerae lipid A secondary hydroxy-acyltransferase and its role in innate immune recognition.
abstract: Similar to most Gram-negative bacteria, the outer leaflet of the outer membrane of Vibrio cholerae is comprised of lipopolysaccharide. Previous reports have proposed that V. cholerae serogroups O1 and O139 synthesize structurally different lipid A domains, which anchor lipopolysaccharide within the outer membrane. In the current study, intact lipid A species of V. cholerae O1 and O139 were analysed by mass spectrometry. We demonstrate that V. cholerae serogroups associated with human disease synthesize a similar asymmetrical hexa-acylated lipid A species, bearing a myristate (C14:0) and 3-hydroxylaurate (3-OH C12:0) at the 2'- and 3'-positions respectively. A previous report from our laboratory characterized the V. cholerae LpxL homologue Vc0213, which transfers a C14:0 to the 2'-position of the glucosamine disaccharide. Our current findings identify V. cholerae Vc0212 as a novel lipid A secondary hydroxy-acyltransferase, termed LpxN, responsible for transferring the 3-hydroxylaurate (3-OH C12:0) to the V. cholerae lipid A domain. Importantly, the presence of a 3-hydroxyl group on the 3'-linked secondary acyl chain was found to promote antimicrobial peptide resistance in V. cholerae; however, this functional group was not required for activation of the innate immune response.
authors.affiliation: Department of Biochemistry and Molecular Biology, Georgia Health Sciences University, Augusta, GA 30912, USA.
authors.forename: Jessica V
authors.initials: JV
authors.lastname: Hankins
authors.forename: James A
authors.initials: JA
authors.lastname: Madsen
authors.forename: David K
authors.initials: DK
authors.lastname: Giles
authors.forename: Brandon M
authors.initials: BM
authors.lastname: Childers
authors.forename: Karl E
authors.initials: KE
authors.lastname: Klose
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Brodbelt
authors.forename: M Stephen
authors.initials: MS
authors.lastname: Trent
mesh_terms: D000217:Acyltransferases / Q000276:immunology*; D002462:Cell Membrane / Q000276:immunology* / Q000648:ultrastructure; D002771:Cholera / Q000276:immunology / Q000382:microbiology; D024881:Drug Resistance, Bacterial; D057809:HEK293 Cells; D006801:Humans; D007113:Immunity, Innate; D008050:Lipid A / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology*; D013058:Mass Spectrometry; D019081:O Antigens / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D011112:Polymyxin B / Q000494:pharmacology; D014734:Vibrio cholerae / Q000201:enzymology / Q000276:immunology*
chemical_list: D008050:Lipid A; D008070:Lipopolysaccharides; D019081:O Antigens; D000217:Acyltransferases; D011112:Polymyxin B