title: Polar Glycosylated and Lateral Non-Glycosylated Flagella from Aeromonas hydrophila Strain AH-1 (Serotype O11).
abstract: Polar and but not lateral flagellin proteins from Aeromonas hydrophila strain AH-1 (serotype O11) were found to be glycosylated. Top-down mass spectrometry studies of purified polar flagellins suggested the presence of a 403 Da glycan of mass. Bottom-up mass spectrometry studies showed the polar flagellin peptides to be modified with 403 Da glycans in O-linkage. The MS fragmentation pattern of this putative glycan was similar to that of pseudaminic acid derivative. Mutants lacking the biosynthesis of pseudaminic acid (pseB and pseI homologues) were unable to produce polar flagella but no changes were observed in lateral flagella by post-transcriptional regulation of the flagellin. Complementation was achieved by reintroduction of the wild-type pseB and pseI. We compared two pathogenic features (adhesion to eukaryotic cells and biofilm production) between the wild-type strain and two kinds of mutants: mutants lacking polar flagella glycosylation and lacking the O11-antigen lipopolysaccharide (LPS) but with unaltered polar flagella glycosylation. Results suggest that polar flagella glycosylation is extremely important for A. hydrophila AH-1 adhesion to Hep-2 cells and biofilm formation. In addition, we show the importance of the polar flagella glycosylation for immune stimulation of IL-8 production via toll-"like" receptor 5 (TLR5).
authors.affiliation: National Research Council, 100 Sussex Drive, Ottawa, ON K1A0R1, Canada. Kelly.Fulton@nrc-cnrc.gc.ca.
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Fulton
authors.affiliation: Departamento de Microbiología, Facultad de Biología, Universidad de Barcelona, Diagonal 645, Barcelona 08071, Spain. elenademendoza@hotmail.com.
authors.forename: Elena
authors.initials: E
authors.lastname: Mendoza-Barberá
authors.affiliation: National Research Council, 100 Sussex Drive, Ottawa, ON K1A0R1, Canada. Susan.Twine@nrc-cnrc.gc.ca.
authors.forename: Susan M
authors.initials: SM
authors.lastname: Twine
authors.affiliation: Departamento de Microbiología, Facultad de Biología, Universidad de Barcelona, Diagonal 645, Barcelona 08071, Spain. jtomas@ub.edu.
authors.forename: Juan M
authors.initials: JM
authors.lastname: Tomás
authors.affiliation: Departamento de Microbiología, Facultad de Biología, Universidad de Barcelona, Diagonal 645, Barcelona 08071, Spain. smerino@ub.edu.
authors.forename: Susana
authors.initials: S
authors.lastname: Merino
mesh_terms: D016980:Aeromonas hydrophila / Q000145:classification / Q000378:metabolism* / Q000648:ultrastructure; D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D018441:Biofilms; D002460:Cell Line; D005407:Flagella / Q000378:metabolism*; D005408:Flagellin / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D059748:Proteolysis; D065288:Serogroup
chemical_list: D005408:Flagellin
keywords: O-flagellin polar glycosylation; adhesion; biofilm; immune stimulation; lateral flagellin non-glycosylated