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Theoretical and experimental characterization of the scope of protein O-glycosylation in Bacteroides fragilis.
- Source :
-
The Journal of biological chemistry [J Biol Chem] 2011 Feb 04; Vol. 286 (5), pp. 3219-26. Date of Electronic Publication: 2010 Nov 29. - Publication Year :
- 2011
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Abstract
- Among bacterial species demonstrated to have protein O-glycosylation systems, that of Bacteroides fragilis and related species is unique in that extracytoplasmic proteins are glycosylated at serine or threonine residues within the specific three-amino acid motif D(S/T)(A/I/L/M/T/V). This feature allows for computational analysis of the proteome to identify candidate glycoproteins. With the criteria of a signal peptidase I or II cleavage site or a predicted transmembrane-spanning region and the presence of at least one glycosylation motif, we identified 1021 candidate glycoproteins of B. fragilis. In addition to the eight glycoproteins identified previously, we confirmed that another 12 candidate glycoproteins are in fact glycosylated. These included four glycoproteins that are predicted to localize to the inner membrane, a compartment not previously shown to include glycosylated proteins. In addition, we show that four proteins involved in cell division and chromosomal segregation, two of which are encoded by candidate essential genes, are glycosylated. To date, we have not identified any extracytoplasmic proteins containing a glycosylation motif that are not glycosylated. Therefore, based on the list of 1021 candidate glycoproteins, it is likely that hundreds of proteins, comprising more than half of the extracytoplasmic proteins of B. fragilis, are glycosylated. Site-directed mutagenesis of several glycoproteins demonstrated that all are glycosylated at the identified glycosylation motif. By engineering glycosylation motifs into a naturally unglycosylated protein, we are able to bring about site-specific glycosylation at the engineered sites, suggesting that this glycosylation system may have applications for glycoengineering.
- Subjects :
- Amino Acid Motifs
Amino Acid Sequence
Aspartic Acid Endopeptidases
Bacterial Proteins
Bacteroides fragilis metabolism
Binding Sites
Membrane Proteins
Mutagenesis, Site-Directed
Serine Endopeptidases
Bacteroides fragilis chemistry
Computational Biology methods
Conserved Sequence
Glycosylation
Proteome
Subjects
Details
- Language :
- English
- ISSN :
- 1083-351X
- Volume :
- 286
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- The Journal of biological chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 21115495
- Full Text :
- https://doi.org/10.1074/jbc.M110.194506