Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Brucella abortus [ICD11:
XN7A8 
]
NCBI PubMed ID: 9006923Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Institutions: Animal Diseases Research Institute, Nepean, ON, Canada
An antigenic O-chain polysaccharide fragment derived from Brucella abortus lipopolysaccharide was labeled with 14.8 +/- 1.8 (n = 5) and 52.3 +/- 2.4 (n = 3) micromol of fluorescein/g of polysaccharide (designated FL1 and FL2, respectively) for use in investigating the binding of O-chain to a specific murine antibody YsT9 under equilibrium conditions. Upon binding to YsT9, the fluorescence of FL1 and FL2 was quenched 45-57% with no shift in the excitation and emission spectra, and polarization of fluorescence increased by 300-335%. With fluorescence quenching and polarization as sensitive signals for antibody-bound labeled O-chains, the equilibrium constants for binding of FL1, FL2, and unlabeled O-chain to YsT9 were determined to be within a similar order (1.5 x 10(7) to 2.0 x 10(7) M-1) using a nonlinear curve fitting approach rather than Scatchard analysis. These results indicated that covalent attachment of fluorescein groups to the O-chain did not influence the recognition of the YsT9-defined epitope by the antibody. The reversibility of the O-chain-antibody reaction was also demonstrated by showing a rapid depolarization of the labeled O-chain-antibody complex in the presence of unlabeled O-chain, suggesting that this displacement experiment could be exploited to quantify the Brucella polysaccharide antigen. The study described here provides a useful model for characterization of the complex formation between a carbohydrate-binding protein and a carbohydrate ligand and also for the design of a homogeneous assay system to quantitate antigens or antibodies of clinical interest.
Lipopolysaccharide, monoclonal antibodies, Brucella, Brucella abortus, binding, fluorescence, polarization, quantitative
Structure type: homopolymer
Location inside paper: p.2821
Trivial name: perosamine homopolymer, perosamine
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_131172,IEDB_134281,IEDB_1397515,IEDB_2116320,IEDB_434547,IEDB_628715
Biological activity: Binding to a monoclonal antibody
Comments, role: Yersinia enterocolitica O9 has the same O-polysaccharide
Related record ID(s): 220, 1236, 3181, 5252, 6859, 7225, 10088, 108433, 110473, 122777, 130189, 143514, 148072
NCBI Taxonomy refs (TaxIDs): 235Reference(s) to other database(s): GTC:G55700WU, GlycomeDB:
3393, CCSD:
42651, CBank-STR:4424
Show glycosyltransferases
There is only one chemically distinct structure:
Found 
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