Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1980 Jul;143(1):328–337. doi: 10.1128/jb.143.1.328-337.1980

Cross-reactivity of major outer membrane proteins of Enterobacteriaceae, studied by crossed immunoelectrophoresis.

H Hofstra, J D Van Tol, J Dankert
PMCID: PMC294238  PMID: 6995435

Abstract

Outer membrane fractions were prepared from 11 bacteria in the family Enterobacteriaceae: Escherichia coli serotypes O1K-, O4K2, O26K60, O75K-, and O111K58, Shigella flexneri, Salmonella typhimurium, Klebsiella pneumonia, Serratia marcescens, Proteus vulgaris, Proteus mirabilis, and Providencia stuartii. All strains studied were found to contain one non-peptidoglycan-bound, heat-modifiable outer membrane protein, and one or two peptidoglycan-associated major outer membrane proteins in the 27,000- to 40,000-dalton range. Crossed immunoelectrophoresis using sodium dodecyl sulfate-polyacarylamide gel electrophoresis for separation of the antigens in the first dimension of the procedure was shown to provide a useful model system for studying the antigenic relationships of the major outer membrane proteins in Enterobacteriaceae species. Peptidoglycan-bound major outer membrane proteins of all bacteria studied reacted with antiserum against the purified peptidogylcan-bound matrix protein I of E. coli O26K60 in this system. Non-peptidoglycan-associated proteins of all strains cross-reacted with protein II of E. coli O26K60 in both their unmodified and their heat-modified forms. These results indicate that the genes coding for the major outer membrane proteins in the family Enterobacteriaceae have been well enough conserved during the course of evolution to allow significant antigenic cross-reactivity between the corresponding proteins in different enterobacterial species.

Full text

PDF
328

Images in this article

331Image
on p.331
332Image
on p.332
334Image
on p.334
335Image
on p.335

Selected References

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

  1. Ames G. F. Resolution of bacterial proteins by polyacrylamide gel electrophoresis on slabs. Membrane, soluble, and periplasmic fractions. J Biol Chem. 1974 Jan 25;249(2):634–644. [PubMed] [Google Scholar]
  2. Bassford P. J., Jr, Diedrich D. L., Schnaitman C. L., Reeves P. Outer membrane proteins of Escherichia coli. VI. Protein alteration in bacteriophage-resistant mutants. J Bacteriol. 1977 Aug;131(2):608–622. doi: 10.1128/jb.131.2.608-622.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Braun V., Rehn K. Chemical characterization, spatial distribution and function of a lipoprotein (murein-lipoprotein) of the E. coli cell wall. The specific effect of trypsin on the membrane structure. Eur J Biochem. 1969 Oct;10(3):426–438. doi: 10.1111/j.1432-1033.1969.tb00707.x. [DOI] [PubMed] [Google Scholar]
  4. Converse C. A., Papermaster D. S. Membrane protein analysis by two-dimensional immunoelectrophoresis. Science. 1975 Aug 8;189(4201):469–472. doi: 10.1126/science.1154021. [DOI] [PubMed] [Google Scholar]
  5. Dankert J., Hofstra H. Antibodies against outer membrane proteins in rabbit antisera prepared against Escherichia coli O26 K60. J Gen Microbiol. 1978 Feb;104(2):311–320. doi: 10.1099/00221287-104-2-311. [DOI] [PubMed] [Google Scholar]
  6. Datta D. B., Krämer C., Henning U. Diploidy for a structural gene specifying a major protein of the outer cell envelope membrane from Escherichia coli K-12. J Bacteriol. 1976 Dec;128(3):834–841. doi: 10.1128/jb.128.3.834-841.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Foulds J., Chai T. J. New major outer membrane proteins found in an Escherichia coli tolF mutant resistant to bacteriophage TuIb. J Bacteriol. 1978 Mar;133(3):1478–1483. doi: 10.1128/jb.133.3.1478-1483.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Galanos C., Lüderitz O., Westphal O. Preparation and properties of antisera against the lipid-A component of bacterial lipopolysaccharides. Eur J Biochem. 1971 Dec 22;24(1):116–122. doi: 10.1111/j.1432-1033.1971.tb19661.x. [DOI] [PubMed] [Google Scholar]
  9. Garten W., Hindennach I., Henning U. The major proteins of the Escherichia coli outer cell envelope membrane. Characterization of proteins II* and III, comparison of all proteins. Eur J Biochem. 1975 Nov 1;59(1):215–221. doi: 10.1111/j.1432-1033.1975.tb02444.x. [DOI] [PubMed] [Google Scholar]
  10. Henning U., Schmidmayr W., Hindennach I. Major proteins of the outer cell envelope membrane of Escherichia coli K-12: multiple species of protein I. Mol Gen Genet. 1977 Sep 9;154(3):293–298. doi: 10.1007/BF00571285. [DOI] [PubMed] [Google Scholar]
  11. Hoff G. E., Frasch C. E. Outer membrane antigens of Neisseria meningitidis group B serotype 2 studied by crossed immunoelectrophoresis. Infect Immun. 1979 Sep;25(3):849–856. doi: 10.1128/iai.25.3.849-856.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hofstra H., Dankert J. Antigenic cross-reactivity of major outer membrane proteins in enterobacteriaceae species. J Gen Microbiol. 1979 Apr;111(2):293–302. doi: 10.1099/00221287-111-2-293. [DOI] [PubMed] [Google Scholar]
  13. Hofstra H., Dankert J. Preparation and quantitative determination of antibodies against major outer mambranes proteins of Escherichia coli O26 K60. J Gen Microbiol. 1980 Apr;117(2):437–447. doi: 10.1099/00221287-117-2-437. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Lee D. R., Schnaitman C. A., Pugsley A. P. Chemical heterogeneity of major outer membrane pore proteins of Escherichia coli. J Bacteriol. 1979 Jun;138(3):861–870. doi: 10.1128/jb.138.3.861-870.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lugtenberg B., Bronstein H., van Selm N., Peters R. Peptidoglycan-associated outer membrane proteins in gammegatine bacteria. Biochim Biophys Acta. 1977 Mar 17;465(3):571–578. doi: 10.1016/0005-2736(77)90274-7. [DOI] [PubMed] [Google Scholar]
  17. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [PubMed] [Google Scholar]
  18. Lüderitz O., Staub A. M., Westphal O. Immunochemistry of O and R antigens of Salmonella and related Enterobacteriaceae. Bacteriol Rev. 1966 Mar;30(1):192–255. doi: 10.1128/br.30.1.192-255.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nakamura K., Pirtle R. M., Inouye M. Homology of the gene coding for outer membrane lipoprotein within various Gram-negative bacteria. J Bacteriol. 1979 Jan;137(1):595–604. doi: 10.1128/jb.137.1.595-604.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Orskov I., Orskov F., Jann B., Jann K. Serology, chemistry, and genetics of O and K antigens of Escherichia coli. Bacteriol Rev. 1977 Sep;41(3):667–710. doi: 10.1128/br.41.3.667-710.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Owen P., Salton M. R. Antigenic and enzymatic architecture of Micrococcus lysodeikticus membranes established by crossed immunoelectrophoresis. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3711–3715. doi: 10.1073/pnas.72.9.3711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Palva E. T., Randall L. L. Arrangement of protein I in Escherichia coli outer membrane: cross-linking study. J Bacteriol. 1978 Jan;133(1):279–286. doi: 10.1128/jb.133.1.279-286.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pugsley A. P., Schnaitman C. A. Outer membrane proteins of Escherichia coli. VII. Evidence that bacteriophage-directed protein 2 functions as a pore. J Bacteriol. 1978 Mar;133(3):1181–1189. doi: 10.1128/jb.133.3.1181-1189.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Reithmeier R. A., Bragg P. D. Purification and characterization of heat-modifiable protein from the outer membrane of Escherichia coli. FEBS Lett. 1974 May 1;41(2):195–198. doi: 10.1016/0014-5793(74)81210-x. [DOI] [PubMed] [Google Scholar]
  25. Rosenbusch J. P. Characterization of the major envelope protein from Escherichia coli. Regular arrangement on the peptidoglycan and unusual dodecyl sulfate binding. J Biol Chem. 1974 Dec 25;249(24):8019–8029. [PubMed] [Google Scholar]
  26. Schmitges C. J., Henning U. The major proteins of the Escherichia coli outer cell-envelope membrane. Heterogeneity of protein I. Eur J Biochem. 1976 Mar 16;63(1):47–52. doi: 10.1111/j.1432-1033.1976.tb10205.x. [DOI] [PubMed] [Google Scholar]
  27. Schnaitman C. A. Solubilization of the cytoplasmic membrane of Escherichia coli by Triton X-100. J Bacteriol. 1971 Oct;108(1):545–552. doi: 10.1128/jb.108.1.545-552.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Smyth C. J., Friedman-Kien A. E., Salton M. R. Antigenic analysis of Neisseria gonorrhoeae by crossed immunoelectrophoresis. Infect Immun. 1976 Apr;13(4):1273–1288. doi: 10.1128/iai.13.4.1273-1288.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Smyth C. J., Siegel J., Salton M. R., Owen P. Immunochemical analysis of inner and outer membranes of Escherichia coli by crossed immunoelectrophoresis. J Bacteriol. 1978 Jan;133(1):306–319. doi: 10.1128/jb.133.1.306-319.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Verhoef C., Lugtenberg B., van Boxtel R., de Graaff P., Verheij H. Genetics and biochemistry of the peptidoglycan-associated proteins b and c of Escherichia coli K12. Mol Gen Genet. 1979 Jan 31;169(2):137–146. doi: 10.1007/BF00271664. [DOI] [PubMed] [Google Scholar]
  31. van Alphen W., van Seim N., Lugtenberg B. Pores in the outer membrane of Escherichia coli K12: involvement of proteins b and e in the functioning of pores for nucleotides. Mol Gen Genet. 1978 Feb 7;159(1):75–83. doi: 10.1007/BF00401750. [DOI] [PubMed] [Google Scholar]
  32. van Raamsdonk W., Pool C. W., Heyting C. Detection of antigens and antibodies by an immuno-peroxidase method applied on thin longitudinal sections of SDS-polyacrylamide gels. J Immunol Methods. 1977;17(3-4):337–348. doi: 10.1016/0022-1759(77)90116-8. [DOI] [PubMed] [Google Scholar]

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

RESOURCES