Taxonomic group: bacteria / Firmicutes
(Phylum: Firmicutes)
Associated disease: infection due to Clostridium difficile [ICD11:
XN0SE 
]
NCBI PubMed ID: 35065968Publication DOI: 10.1016/j.jbc.2022.101622Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: P.J.Hensbergen
lumc.nl
Institutions: Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands, Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands
The type A glycan modification found in human pathogen Clostridioides difficile consists of a monosaccharide (GlcNAc) that is linked to an N-methylated threonine through a phosphodiester bond. This structure has previously been described on the flagellar protein flagellin C of several C. difficile strains and is important for bacterial motility. The study of post-translational modifications often relies on some type of enrichment strategy; however, a procedure for enrichment of this modification has not yet been demonstrated. In this study, we show that an approach that is commonly used in phosphoproteomics, Fe3+-immobilized metal affinity chromatography, also enriches for peptides with this unique post-translational modification. Using LC-MS/MS analyses of immobilized metal affinity chromatography-captured tryptic peptides, we observed not only type A-modified C. difficile flagellin peptides but also a variety of truncated/modified type A structures on these peptides. Using an elaborate set of mass spectrometry analyses, we demonstrate that one of these modifications consists of a type A structure containing a phosphonate (2-aminoethylphosphonate), a modification that is rarely observed and has hitherto not been described in C. difficile. In conclusion, we show that a common enrichment strategy results in reliable identification of peptides carrying a type A glycan modification, and that the results obtained can be used to advance models about its biosynthesis.
mass spectrometry, glycosylation, IMAC, Clostridioides, FliC, phosphonate
Structure type: monomer
Location inside paper: Fig. 1A, type A modification
Aglycon: Ser/Thr
Trivial name: flagellin C (FliC)
Contained glycoepitopes: IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531
Methods: LC-MS/MS, IMAC, data analysis
NCBI Taxonomy refs (TaxIDs): 272563
Show glycosyltransferases
There is only one chemically distinct structure:
Found 
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(previously named: Clostridium difficile 630Δerm)