Posttranslational modification of flagellin flaB in Shewanella oneidensis

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DOIResolve DOI: http://doi.org/10.1128/JB.00015-13
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TypeArticle
Journal titleJournal of Bacteriology
ISSN0021-9193
Volume195
Issue11
Pages25502561; # of pages: 12
Subjectcysteine; flagellin; lysine; protein FlaB; protein PseB; protein PseC; serine; bacterial strain; bacterium mutant; controlled study; glycosylation; liquid chromatography; methylation; protein isolation; protein processing; Salmonella enterica; Shewanella oneidensis; Amino Acid Sequence; Amino Acid Substitution; Bacterial Proteins; Chromatography, Liquid; Flagella; Genetic Complementation Test; Glycosylation; Mass Spectrometry; Mutagenesis, Site-Directed; Mutation; Polysaccharides; Protein Processing, Post-Translational; Recombinant Fusion Proteins; Tandem Mass Spectrometry
AbstractShewanella oneidensis is a highly motile organism by virtue of a polar, glycosylated flagellum composed of flagellins FlaA and FlaB. In this study, the functional flagellin FlaB was isolated and analyzed with nano-liquid chromatography-mass spectrometry (MS) and tandem MS. In combination with the mutational analysis, we propose that the FlaB flagellin protein from S. oneidensis is modified at five serine residues with a series of novel O-linked posttranslational modifications (PTMs) that differ from each other by 14 Da. These PTMs are composed in part of a 274-Da sugar residue that bears a resemblance to the nonulosonic acids. The remainder appears to be composed of a second residue whose mass varies by 14 Da depending on the PTM. Further investigation revealed that synthesis of the glycans initiates with PseB and PseC, the first two enzymes of the Pse pathway. In addition, a number of lysine residues are found to be methylated by SO4160, an analogue of the lysine methyltransferase of Salmonella enterica serovar Typhimurium.
Publication date
LanguageEnglish
AffiliationHuman Health Therapeutics; National Research Council Canada
Peer reviewedYes
NPARC number21270469
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Record identifier32c95948-d4eb-4871-a7c4-aa51276a066c
Record created2014-02-12
Record modified2016-05-09
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