Structural studies of the Vibrio cholerae o-antigen

Kenne, Lennart; Lindberg, Bengt; Unger, Per; Gustafsson, Björn; Holme, Tord

doi:10.1016/s0008-6215(00)81047-2

Cited by 140 publications

(76 citation statements)

References 15 publications

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“…PCR products were cloned after gel purification and cleavage with HindII and HindIII (Boehringer Mannheim). The complete sequences of the Sac I fragment harboring rib from 017 and 569B were determined by using deoxyadenosine 5 Sequenase (United States Biochemical). Cloned PCR products were sequenced on an Applied Biosystems 373A automated sequencer using both dye primers and terminators.…”

Section: Methodsmentioning

confidence: 99%

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Serotype conversion in Vibrio cholerae O1.

Stroeher

Karageorgos

Morona

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

178

166

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Vibrio cholerae 01 exists as two major serotypes, Inaba and Ogawa, which are associated with the 0 antigen of the lipopolysaccharide and are capable of unequal reciprocal interconversion. The 20-kilobase rJb regions encoding O-antigen biosynthesis in strains 569B (Inaba) and 017 (Ogawa) have been cloned in Escherichia coi K-12 and the nucleotide sequences have been determined. Besides several base substitutions and a small deletion in the 569B sequence relative to 017, there is a single nucleotide change resulting in a TGA stop codon within the gene for the 32-kDa RfbT protein.

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Section: Methodsmentioning

confidence: 99%

“…Analysis of the chemical structure of the 0 antigen has shown it to be composed of a homopolymer containing the amino sugar D-perosamine substituted with 3-deoxy-Lglycerotetronic acid (5,6). This structure is present in both Inaba and Ogawa serotypes and thus may be the A antigen.…”

mentioning

confidence: 99%

Serotype conversion in Vibrio cholerae O1.

Stroeher

Karageorgos

Morona

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

178

166

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“…The O polysaccharide structures of the cholera causing serogroups O1 and O139 (13,14), as well as the underlying core OS, have been resolved (15,16). Compared with O1, the core OS of two non-O1 isolates, H11 (17), and O22 (14,18), revealed distinct structural differences.…”

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confidence: 99%

Molecular and Functional Characterization of O Antigen Transfer inVibriocholerae

Schild

Lamprecht

Reidl

2005

Journal of Biological Chemistry

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The outer lipid leaflet of the outer membrane of Gram-negative bacteria (1) is composed almost exclusively of lipopolysaccharide (LPS) 1 consisting of three complex portions, i.e. the lipid A, the core oligosaccharides (core OS), and the O polysaccharides. As a polyanionic lipid, the LPS leaflet acts as an effective barrier for the diffusion of lipophilic and hydrophobic compounds. This characteristic depends on the lipid A-core OS complex (for a recent summary, see Ref. 2).

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“…We used synthetic mono-to hexasaccharides that mimic the fragments of the O-antigen of Ogawa and Inaba O-polysaccharides (2)(3)(4), together with certain analogs of their monosaccharides to evaluate specificity. The binding of three immunoglobulins G (two specific for Ogawa and one specific for Ogawa/ Inaba) and of two immunoglobulins A (one specific for Ogawa and one specific for Inaba/Ogawa) were characterized by ligand-induced fluorescence titration or ELISA inhibition.…”

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confidence: 99%

On the Antigenic Determinants of the Lipopolysaccharides of Vibrio cholerae O:1, Serotypes Ogawa and Inaba

Wang¹,

Villeneuve²,

Zhang³

et al. 1998

Journal of Biological Chemistry

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We used synthetic mono-to hexasaccharides that mimic the fragments of the O-antigen of Ogawa and Inaba O-polysaccharides (2-4), together with certain analogs of their monosaccharides to evaluate specificity. The binding of three immunoglobulins G (two specific for Ogawa and one specific for Ogawa/ Inaba) and of two immunoglobulins A (one specific for Ogawa and one specific for Inaba/Ogawa) were characterized by ligand-induced fluorescence titration or ELISA inhibition. The cDNA sequences of these antibodies are also presented in this report. MATERIALS AND METHODSMonoclonal Antibodies-Murine ascites fluids of A-20-6 and S-20-4 both contain vibriocidal IgG 1 specific for Ogawa-LPS. I-24-2, in contrast, contains IgG 3 specific for both serotypes Ogawa and Inaba-LPS, and it has low vibriocidal activity (5) (clone S-20-4 comes from the same hybridoma cells as clone S-20-3 described in this reference). Murine ascites fluid 2D6 and ZAC-3 contain IgA specific for Ogawa-LPS and IgA specific for Inaba/Ogawa-LPS, respectively. The latter two hybridomas were gifts from Drs. Marian Neutra, Harvard Medical School, and Dr. Richard Weltzin, Oravax, Cambridge, MA (1, 6) and were grown in BALB/c mice. IgGs were purified using ImmunoPure® (G) IgG purification kits (Pierce). Briefly, ascites fluid (2 ml, clarified by centrifugation) was mixed with ImmunoPure® (G) binding buffer (2 ml) and applied to a protein G column. After washing the column with 5 ϫ 2-ml aliquots of the ImmunoPure® (G) binding buffer, the bound IgG was eluted with 6 ml of ImmunoPure® (G) elution buffer, dialyzed against PBS, pH 7.4 (2000 ml) for three changes at 0°C, frozen, and labeled. The purified A-20-6, S-20-4, and I-24-2 showed a single arc of precipitation versus goat anti-mouse IgG 1 and IgG 3 , respectively (heavy chainspecific), and goat anti-whole mouse serum (Sigma) by immunoelectrophoresis. IgAs were purified from ascites fluid by 40% ammonium sulfate precipitation and anion-exchange DEAE-Sephadex A-25 chromatography (7). Monomeric IgA was obtained by mild reduction with 5 mM 1,4-dithiothreitol (Sigma) and alkylation with 11 mM 2-iodoacetamide (Sigma), followed by re-adsorption of the sample on DEAESephadex A-25 and elution with PBS, pH 7.4. The purity of IgAs was also verified by immuno-electrophoresis against anti-mouse IgA and serum and SDS-polyacrylamide gel electrophoresis.LPS and Synthetic Oligosaccharides-V. cholerae O:1 LPSs were obtained from acetone-treated cells of strain 569B, classical biotype, serotype Inaba, lot VC1219; strain 3083, classical biotype, serotype Ogawa; and V. cholerae O:139 Bengal, strain 4450. Salmonella paratyphi A LPS was a field isolate in Nepal, strain NTP-6. All LPSs were purified as described (8) and at 2 mg/ml showed negative tests (Coomassie Blue) for protein. Severely base-degraded V. cholerae O:139 LPS (9) was a gift from Dr. Andrew D. Cox, National Research Council, Ottawa, Canada. De-O-acylated Ogawa-LPS (10) was oxidized in aqueous 0.8% periodate solution for 3 days in the dark, dialyzed, and freezedried a...

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Structural studies of the Vibrio cholerae o-antigen

Cited by 140 publications

References 15 publications

Serotype conversion in Vibrio cholerae O1.

Serotype conversion in Vibrio cholerae O1.

Molecular and Functional Characterization of O Antigen Transfer inVibriocholerae

On the Antigenic Determinants of the Lipopolysaccharides of Vibrio cholerae O:1, Serotypes Ogawa and Inaba

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