Structural studies of the O-specific polysaccharide of Vibrio cholerae O8 using solvolysis with triflic acid.
The O-specific polysaccharide (OPS) of Vibrio cholerae 08 was isolated by mild acid degradation of the lipopolysaccharide and studied by two-dimensional NMR spectroscopy, including NOESY and heteronuclear multiple-bond correlation (HMBC) experiments. The OPS was found to have a tetrasaccharide repeating unit with the following structure: --> 4)-beta-D-Glcp NAc3NAcylAN-(1 --> 4)-beta-D-Manp NAc3NAcAN-(1 --> 4)-alpha-L-Gulp NAc3NAcA-(1 --> 3) -beta-D-QuipNAc4NAc-(1 --> where QuiNAc4NAc is 2,4-diacetamido-2,4,6-trideoxyglucose, GlcNAc3NAcylAN is 2-acetamido-3-(N-formyl-L-alanyl)amino-2,3-dideoxyglucuronamide, ManNAc3NAcAN is 2,3-diacetamido-2,3-dideoxymannuronamide, and GulNAc3NAcA is 2,3-diacetamido-2,3-dideoxyguluronic acid. The OPS was stable towards acid hydrolysis and solvolysis with anhydrous hydrogen fluoride, but could be cleaved selectively with trifluoromethanesulfonic (triflic) acid by the glycosidic linkages of beta-QuiNAc4NAc and alpha-GulNAc3NAcA. The structures of the oligosaccharides obtained that were elucidated by electrospray ionization (ESI) MS and NMR spectroscopy, confirmed the OPS structure.
Kocharova
N A
NA
Karolinska Institute, Clinical Research Centre, Huddinge University Hospital, Sweden.
Perepelov
A V
AV
Zatonsky
G V
GV
Shashkov
A S
AS
Knirel
Y A
YA
Jansson
P E
PE
Weintraub
A
A
D000942:Antigens, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D008698:Mesylates; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D019081:O Antigens / Q000737:chemistry* / Q000302:isolation & purification; D011135:Polysaccharides, Bacterial / Q000276:immunology; D012997:Solvents; D021241:Spectrometry, Mass, Electrospray Ionization; D014734:Vibrio cholerae / Q000276:immunology*
D000942:Antigens, Bacterial; D008698:Mesylates; D019081:O Antigens; D011135:Polysaccharides, Bacterial; D012997:Solvents; C012077:trifluoromethanesulfonic acid