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The S-layer protein of a Clostridium difficile SLCT-11 strain displays a complex glycan required for normal cell growth and morphology.

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Title: The S-layer protein of a Clostridium difficile SLCT-11 strain displays a complex glycan required for normal cell growth and morphology.
Authors: Richards, E
Bouché, L
Panico, M
Arbeloa, A
Vinogradov, E
Morris, H
Wren, B
Logan, SM
Dell, A
Fairweather, NF
Item Type: Journal Article
Abstract: 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. difficilestrain 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 mass spectrometry 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 non-reducing 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. difficilesporulation and control of cell shape, a discovery with implications for the development of antimicrobials targeting the S-layer.
Issue Date: 23-Nov-2018
Date of Acceptance: 1-Oct-2018
URI: http://hdl.handle.net/10044/1/65604
DOI: https://dx.doi.org/10.1074/jbc.RA118.004530
ISSN: 0021-9258
Publisher: American Society for Biochemistry and Molecular Biology
Start Page: 18123
End Page: 18137
Journal / Book Title: Journal of Biological Chemistry
Volume: 293
Copyright Statement: © 2018 Richards et al. Final version open access under the terms of the Creative Commons CC-BY license (https://creativecommons.org/licenses/by/4.0/)
Keywords: Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
mass spectrometry (MS)
nuclear magnetic resonance (NMR)
microbiology
glycoprotein structure
genetics
Clostridium difficile
surface layer
virulence
glycosylation
glycomics
glycoproteomics
cell wall
SURFACE-LAYER
MASS-SPECTROMETRY
ANTHRACIS
GENE
RECONSTRUCTION
GLYCOSYLATION
TRANSMISSION
FLAGELLIN
DOMAINS
06 Biological Sciences
11 Medical And Health Sciences
03 Chemical Sciences
Publication Status: Published
Conference Place: United States
Open Access location: http://www.jbc.org/content/early/2018/10/02/jbc.RA118.004530.1.full.pdf?with-ds=yes
Online Publication Date: 2018-10-01
Appears in Collections:Faculty of Natural Sciences