| Download | - View final version: The type B flagellin of hypervirulent clostridium difficile is modified with novel sulphonated Peptidylamido-glycans (PDF, 1.6 MiB)
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| DOI | Resolve DOI: https://doi.org/10.1074/jbc.M116.749481 |
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| Author | Search for: Bouché, Laura; Search for: Panico, Maria; Search for: Hitchen, Paul; Search for: Binet, Daniel; Search for: Sastre, Federico; Search for: Faulds-pain, Alexandra; Search for: Valiente, Esmeralda; Search for: Vinogradov, Evgeny1; Search for: Aubry, Annie1; Search for: Fulton, Kelly1; Search for: Twine, Susan1; Search for: Logan, Susan m1; Search for: Wren, Brendan w; Search for: Dell, Anne; Search for: Morris, Howard r |
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| Name affiliation | - National Research Council of Canada. Human Health Therapeutics
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| Format | Text, Article |
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| Subject | bacteria; glycosylation; gram-positive bacteria; mass spectrometry (MS); nuclear magnetic resonance (NMR); Clostridium difficile; flagellin; modification; sulphonated |
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| Abstract | Glycosylation of flagellins is a well recognized property of many bacterial species. In this study we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile. We used mass spectrometry (nano LC- MS and MS/MS analysis) to identify a number of putative glycopeptides which carried a variety of glycoform substitutions each of which was linked through an initial HexNAc residue to Ser or Thr. Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. A variety of Electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive- and negative-ion collisionally activated decomposition (CAD) MS/MS which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a Taurine-containing peptidylamido-glycan structure. Finally NMR analysis of flagellin glycopeptides provided complementary information. The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser and the novel capping moiety was shown to be comprised of Taurine, Alanine, and Glycine. This is the first report of a novel O-linked sulphonated peptidylamido-glycan moiety decorating a flagellin protein. |
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| Publication date | 2016-10 |
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| Publisher | American Society for Biochemistry and Molecular Biology |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NPARC number | 23000836 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | 5ac353b2-654a-48eb-9fc4-c05458f7459d |
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| Record created | 2016-10-17 |
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| Record modified | 2020-06-02 |
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