Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1985 Oct;164(1):263–269. doi: 10.1128/jb.164.1.263-269.1985

Electrophoretic and immunochemical analyses of the lipopolysaccharides from various strains of Aeromonas hydrophila.

J S Dooley, R Lallier, D H Shaw, T J Trust
PMCID: PMC214238  PMID: 2413008

Abstract

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to analyze the lipopolysaccharides isolated from strains of Aeromonas hydrophila which exhibit virulence for fish and which autoaggregate during growth in static broth culture. The lipopolysaccharides contained O-polysaccharide chains of homogeneous chain length. Two of the strains produced a surface protein array, and immunofluorescence and phage-binding studies revealed that a number of these O-polysaccharide chains of homogeneous length traversed the protein array and were exposed on the cell surface. Immunochemical analyses by immunoblotting, enzyme-linked immunosorbent assay, immunofluorescence, and immunoprecipitation with both polyclonal and monoclonal antibodies revealed the presence of three epitopes on the polysaccharide moiety of this homogenous-chain-length lipopolysaccharide morphotype. One epitope was species serogroup specific and reactive by immunoblotting. This epitope was not present on the heterogeneous-chain-length O polysaccharides of nonautoaggregating strains of A. hydrophila examined. The second epitope was conformation dependent and cross-reactive with an epitope on the homogenous-chain-length O polysaccharides of Aeromonas salmonicida lipopolysaccharide. The third epitope was recognized by a monoclonal antibody and appeared to involve that region of the A. hydrophila and A. salmonicida lipopolysaccharide molecules which contained the O-polysaccharide-core oligosaccharide glycosidic linkage.

Full text

PDF
268

Images in this article

265Image
on p.265
265Image
on p.265
266Image
on p.266
267Image
on p.267
267Image
on p.267

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chart H., Shaw D. H., Ishiguro E. E., Trust T. J. Structural and immunochemical homogeneity of Aeromonas salmonicida lipopolysaccharide. J Bacteriol. 1984 Apr;158(1):16–22. doi: 10.1128/jb.158.1.16-22.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. J Immunol. 1972 Jul;109(1):129–135. [PubMed] [Google Scholar]
  3. Goldman R. C., Leive L. Heterogeneity of antigenic-side-chain length in lipopolysaccharide from Escherichia coli 0111 and Salmonella typhimurium LT2. Eur J Biochem. 1980;107(1):145–153. doi: 10.1111/j.1432-1033.1980.tb04635.x. [DOI] [PubMed] [Google Scholar]
  4. Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ishiguro E. E., Ainsworth T., Shaw D. H., Kay W. W., Trust T. J. A lipopolysaccharide-specific bacteriophage for Aeromonas salmonicida. Can J Microbiol. 1983 Oct;29(10):1458–1461. doi: 10.1139/m83-223. [DOI] [PubMed] [Google Scholar]
  6. Jann B., Reske K., Jann K. Heterogeneity of lipopolysaccharides. Analysis of polysaccharide chain lengths by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Eur J Biochem. 1975 Dec 1;60(1):239–246. doi: 10.1111/j.1432-1033.1975.tb20996.x. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Logan S. M., Trust T. J. Structural and antigenic heterogeneity of lipopolysaccharides of Campylobacter jejuni and Campylobacter coli. Infect Immun. 1984 Jul;45(1):210–216. doi: 10.1128/iai.45.1.210-216.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mittal K. R., Lalonde G., Leblanc D., Olivier G., Lallier R. Aeromonas hydrophila in rainbow trout: relation between virulence and surface characteristics. Can J Microbiol. 1980 Dec;26(12):1501–1503. doi: 10.1139/m80-248. [DOI] [PubMed] [Google Scholar]
  10. Palva E. T., Mäkelä P. H. Lipopolysaccharide heterogeneity in Salmonella typhimurium analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Eur J Biochem. 1980;107(1):137–143. doi: 10.1111/j.1432-1033.1980.tb04634.x. [DOI] [PubMed] [Google Scholar]
  11. Pearson T. W., Pinder M., Roelants G. E., Kar S. K., Lundin L. B., Mayor-Withey K. S., Hwett R. S. Methods for derivation and detection of anti-parasite monoclonal antibodies. J Immunol Methods. 1980;34(2):141–154. doi: 10.1016/0022-1759(80)90168-4. [DOI] [PubMed] [Google Scholar]
  12. Shaw D. H., Lee Y. Z., Squires M. J., Lüderitz O. Structural studies on the O-antigen of Aeromonas salmonicida. Eur J Biochem. 1983 Apr 5;131(3):633–638. doi: 10.1111/j.1432-1033.1983.tb07310.x. [DOI] [PubMed] [Google Scholar]
  13. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES