Taxonomic group: bacteria / Firmicutes
(Phylum: Firmicutes)
Associated disease: infection due to Bacillus anthracis [ICD11:
XN94F 
]
NCBI PubMed ID: 22012665Publication DOI: 10.1002/jms.1980Journal NLM ID: 9504818Publisher: Chichester, UK: Wiley
Correspondence: banoubjo
dfo-mpo.gc.ca
Institutions: Department of Chemistry, Memorial University of Newfoundland, Saint John's, NL, Canada
We present herein an efficient mass spectrometric method for the localization of the glycation sites of a model neoglycoconjugate vaccine formed by a construct of the tetrasaccharide side chain of the Bacillus anthracis exosporium and the protein carrier bovine serum albumin. The glycoconjugate was digested with both trypsin and GluC V8 endoproteinases, and the digests were then analyzed by MALDI-TOF/TOF-CID-MS/MS and nano-LC-ESI-QqTOF-CID-MS/MS. The sequences of the unknown peptides analyzed by MALDI-TOF/TOF-CID-MS/MS, following digestion with the GluC V8 endoproteinase, allowed us to recognize three glycopeptides whose glycation occupancies were, respectively, on Lys 235, Lys 420, and Lys 498. Similarly, the same analysis was performed on the tryptic digests, which permitted us to recognize two glycation sites on Lys 100 and Lys 374. In addition, we have also used LC-ESI-QqTOF-CID-MS/MS analysis for the identification of the tryptic digests. However, this analysis identified a higher number of glycopeptides than would be expected from a glycoconjugate composed of a carbohydrate-protein ratio of 5.4:1, which would have resulted in glycation occupancies of 18 specific sites. This discrepancy was due to the large number of glycoforms formed during the synthetic carbohydrate-spacer-carrier protein conjugation. Likewise, the LC-ESI-QqTOF-MS/MS analysis of the GluC V8 digest also identified 17 different glycation sites on the synthetic glycoconjugate.
neoglycoconjugate, proteinases, glycated peptides, MALDI-TOF/TOF-CID-MS/MS, LC-ESI-QqTOF-CID-MS/MS
Structure type: oligomer
Location inside paper: p.994, fig.1
Aglycon: spacer-BSA
Contained glycoepitopes: IEDB_133754,IEDB_136105,IEDB_225177,IEDB_885823
Methods: MALDI-TOF/TOF MS, MALDI-CID-TOF/TOF MS/MS, LC-ESI-QTOF MS/MS
Comments, role: synthetic tetrasaccharide
NCBI Taxonomy refs (TaxIDs): 1392
Show glycosyltransferases
There is only one chemically distinct structure:
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