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Taxonomic group: bacteria / Firmicutes (Phylum: Firmicutes)
Host organism: Homo sapiens
Associated disease: functional gastrointestinal disorders [ICD11: DD9Y

]; infection due to Clostridium difficile [ICD11: XN0SE

]

The structure was elucidated in this paper
NCBI PubMed ID: 30275012
Publication DOI: 10.1074/jbc.RA118.004530
Journal NLM ID: 2985121R
Publisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: a.dell

imperial.ac.uk; n.fairweather

imperial.ac.uk
Institutions: From the Department of Life Sciences, Imperial College London, SW7 2AZ London, United Kingdom, the Vaccine Program, Human Health Therapeutics Research Centre, National Research Council, Ottawa, Ontario K1A 0R6, Canada, Biopharmaspec, Suite 3.1, Lido Medical Centre, St. Saviours Road, JE2 7LA Jersey, United Kingdom, the London School of Hygiene and Tropical Medicine, WC1E 7HT, London, United Kingdom

Clostridium difficile is a bacterial pathogen that causes major health challenges worldwide. It has a well-characterized surface (S)-layer, a para-crystalline proteinaceous layer surrounding the cell wall. In many bacterial and archaeal species, the S-layer is glycosylated, but no such modifications have been demonstrated in C. difficile. Here, we show that a C. difficile strain of S-layer cassette type 11, Ox247, has a complex glycan attached via an O-linkage to Thr-38 of the S-layer low-molecular-weight subunit. Using MS and NMR, we fully characterized this glycan. We present evidence that it is composed of three domains: (i) a core peptide-linked tetrasaccharide with the sequence -4-α-Rha-3-α-Rha-3-α-Rha-3-β-Gal-peptide; (ii) a repeating pentasaccharide with the sequence -4-β-Rha-4-α-Glc-3-β-Rha-4-(α-Rib-3-)β-Rha-; and (iii) a nonreducing end-terminal 2,3 cyclophosphoryl-rhamnose attached to a ribose-branched sub-terminal rhamnose residue. The Ox247 genome contains a 24-kb locus containing genes for synthesis and protein attachment of this glycan. Mutations in genes within this locus altered or completely abrogated formation of this glycan, and their phenotypes suggested that this S-layer modification may affect sporulation, cell length, and biofilm formation of C. difficile In summary, our findings indicate that the S-layer protein of SLCT-11 strains displays a complex glycan and suggest that this glycan is required for C. difficile sporulation and control of cell shape, a discovery with implications for the development of antimicrobials targeting the S-layer.

genetics, microbiology, virulence, cell wall, glycosylation, surface layer, Glycomics, Clostridium difficile, nuclear magnetic resonance (NMR), mass spectrometry (MS), Glycoproteomics, glycoprotein structure

Structure type: oligomer
Location inside paper: fig.9
Aglycon: tetrapeptide (Thr38, Thr, Gly, Ala)
Trivial name: S-layer glycan
Compound class: surface polysaccharide
Contained glycoepitopes: IEDB_136044,IEDB_137472,IEDB_1394181,IEDB_141794,IEDB_142488,IEDB_144998,IEDB_145010,IEDB_146664,IEDB_149136,IEDB_190606,IEDB_983931,SB_165,SB_166,SB_187,SB_192,SB_195,SB_7,SB_88

Methods: 13C NMR, 1H NMR, gel filtration, NMR-2D, GC-MS, SDS-PAGE, sugar analysis, DNA techniques, anion-exchange chromatography, MS/MS, LC-ESI-MS, MALDI-TOF/TOF MS, biofilm assays, bioinformatics analysis, adhesion assays, sporulation assays
Biosynthesis and genetic data: Orf19, Orf3, Orf4, Orf7
Comments, role: Proposed overall structure of the glycan attached to SlpA in Ox247.

Related record ID(s): 12967
NCBI Taxonomy refs (TaxIDs): 1496
Reference(s) to other database(s): GTC:G75771PL
Show glycosyltransferases

There is only one chemically distinct structure:

C[C@H]1O[C@@H](O[C@@H]2[C@@H](CO)O[C@H](O[C@@H]3[C@H](O)[C@H](O[C@@H]4[C@@H](C)O[C@@H](O[C@@H]5[C@@H](C)O[C@@H](O[C@@H]6[C@@H](CO)O[C@H](O[C@@H]7[C@H](O)[C@H](O[C@@H]8[C@@H](C)O[C@@H](O[C@@H]9[C@@H](C)O[C@@H](O[C@@H]%10[C@@H](CO)O[C@H](O[C@@H]%11[C@H](O)[C@H](O[C@@H]%12[C@@H](C)O[C@@H](O[C@@H]%13[C@@H](C)O[C@H](O[C@@H]%14[C@H](O)[C@@H](O[C@@H]%15[C@H](O)[C@@H](O[C@H]%16[C@@H](O)[C@@H](CO)O[C@@H](O)[C@@H]%16O)O[C@H](C)[C@H]%15O)O[C@H](C)[C@H]%14O)[C@@H](O)[C@H]%13O)[C@@H](O)[C@H]%12O[C@H]%12O[C@H](CO)[C@@H](O)[C@H]%12O)O[C@H](C)[C@H]%11O)[C@H](O)[C@H]%10O)[C@@H](O)[C@H]9O)[C@@H](O)[C@H]8O[C@H]8O[C@H](CO)[C@@H](O)[C@H]8O)O[C@H](C)[C@H]7O)[C@H](O)[C@H]6O)[C@@H](O)[C@H]5O)[C@@H](O)[C@H]4O[C@H]4O[C@H](CO)[C@@H](O)[C@H]4O)O[C@H](C)[C@H]3O)[C@H](O)[C@H]2O)[C@@H](O)[C@@H](O)[C@@H]1O[C@@H]1O[C@H](C)[C@@H](O[C@@H]2O[C@H](C)[C@@H](O)[C@H](O[C@H]3O[C@H](CO)[C@@H](O[C@@H]4O[C@H](C)[C@@H](O[C@@H]5O[C@H](C)[C@@H](O[C@@H]6O[C@H](C)[C@@H](O)[C@H](O[C@H]7O[C@H](CO)[C@@H](O[C@@H]8O[C@H](C)[C@@H](O[C@@H]9O[C@H](C)[C@@H](O[C@@H]%10O[C@H](C)[C@@H](O)[C@@H]%11OP(=O)(O)O[C@H]%10%11)[C@H](O[C@H]%10O[C@H](CO)[C@@H](O)[C@H]%10O)[C@@H]9O)[C@H](O)[C@@H]8O)[C@H](O)[C@H]7O)[C@@H]6O)[C@H](O[C@H]6O[C@H](CO)[C@@H](O)[C@H]6O)[C@@H]5O)[C@H](O)[C@@H]4O)[C@H](O)[C@H]3O)[C@@H]2O)[C@H](O[C@H]2O[C@H](CO)[C@@H](O)[C@H]2O)[C@@H]1O

SVG MW SMILES 3D mol Ionize

Warning = sub-repeat ending at bDRhap: large unit count (8) was reduced to 5