1887

Abstract

Antisera against porin oligomers isolated from the outer membrane of O26 K60 and against porin monomers from the same bacterial strain were elicited in rabbits by intramuscular administration with Freund’s complete adjuvant. Antibodies against native porin oligomers reacted strongly with porin oligomers, as revealed by sodium dodecyl sulphate-polyacrylamide gel immunoperoxidase (SGIP) analysis, the enzyme-linked immunosorbent assay (ELISA) and immunodiffusion, but showed no significant reaction with denatured monomers. The antibodies were completely absorbed by the intact outer membrane-peptidoglycan complex, which suggests that they were directed against antigenic determinants expressed on the outside of the intact outer membrane. Antibodies directed against denatured porin monomers reacted strongly with monomers in all tests but reacted only very weakly with porin oligomers. They were not absorbed by the native porin situated in the intact outer membrane. This indicates that the major antigenic determinants of the denatured porin monomer are hardly related to those of the native trimer situated in the intact outer membrane. The antigenic determinants of the denatured monomer seem to become fully expressed only after dissociation and denaturation of the porin. It is concluded that the immunological relationship of denatured porin monomers derived from many strains of and other which was reported in previous studies may not indicate that native porin trimers of these strains are also related.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-125-2-285
1981-08-01
2021-05-12
Loading full text...

Full text loading...

/deliver/fulltext/micro/125/2/mic-125-2-285.html?itemId=/content/journal/micro/10.1099/00221287-125-2-285&mimeType=html&fmt=ahah

References

  1. Benz R., Janko K., Boos W., Laüger P. 1978; Formation of large, ion-permeable membrane chan-nels by the matrix protein (porin) of Escherichia coli. Biochimica et biophysica acta 511:305–319
    [Google Scholar]
  2. Dankert J., Hofstra H. 1978; Antibodies against outer membrane proteins in rabbit antisera prepared against Escherichia coli O26K60. Journal of General Microbiology 104:311–320
    [Google Scholar]
  3. Datta D. B., Arden B., Henning U. 1977; Major proteins of the Escherichia coli outer cell envelope membrane as bacteriophage receptors. Journal of Bacteriology 131:821–829
    [Google Scholar]
  4. Decad G. M., Nikaido H. 1976; Outer membrane of Gram-negative bacteria. XII. Molecular sieving function of cell wall. Journal of Bacteriology 128:325–336
    [Google Scholar]
  5. Hasegawa Y., Yamada H., Mizushima S. 1976; Interactions of outer membrane proteins O-8 and O-9 with peptidoglycan sacculus of Escherichia coli K-12. Journal of Biochemistry 80:1401–1409
    [Google Scholar]
  6. Hofstra H., Dankert J. 1979; Antigenic cross-reactivity of major outer membrane proteins in Enterobacteriaceae species. Journal of General Microbiology 111:293–302
    [Google Scholar]
  7. Hofstra H., Dankert J. 1980a; Preparation and quantitative determination of antibodies against major outer membrane proteins of Escherichia coli O26K60. Journal of General Microbiology 117:437–447
    [Google Scholar]
  8. Hofstra H., Dankert J. 1980b; Antigenic cross-reactivity of outer membrane proteins of Escherichia coli and Proteus species. FEMS Microbiology Letters 7:171–174
    [Google Scholar]
  9. Hofstra H., Dankert J. 1980c; Major outer membrane proteins: common antigens in Entero-bacteriaceae species. Journal of General Microbiology 119:123–131
    [Google Scholar]
  10. Hofstra H., Van Tol M. J. D., Dankert J. 1980; Antigenic cross-reactivity of the major outer membrane proteins of Enterobacteriaceae studied by crossed immunoelectrophoresis. Journal of Bacteriology 143:328–337
    [Google Scholar]
  11. Inouye S., Takeishi K., Lee N., Demartini M., Harashima A., Inouye M. 1976; Lipoprotein from the outer membrane of Escherichia coli: purification, paracrystallization and some properties of its free form. Journal of Bacteriology 127:555–563
    [Google Scholar]
  12. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  13. Lugtenberg B., Meuers J., Peters R., Van Der hoek P., Van Alphen L. 1975; Electro-phoretic resolution of the ‘Major outer membrane protein’ of Escherichia coli K12 into four bands. FEBS Letters 58:254–258
    [Google Scholar]
  14. Nakae T. 1976a; Identification of the outer mem-brane protein of Escherichia coli that produces transmembrane channels in reconstituted vesicle membranes. Biochemical and Biophysical Research Communications 71:877–884
    [Google Scholar]
  15. Nakae T. 1976b; Outer membrane of Salmonella.Isolation of protein complex that produces transmembrane channels. Journal of Biological Chem-istry 251:2176–2178
    [Google Scholar]
  16. Nakae T., Nikaido H. 1975; Outer membrane as a diffusion barrier in Salmonella typhimurium: penetration of oligo- and polysaccharides into isolated outer membrane vesicles and cells with degraded peptidoglycan layer. Journal of Biological Chemistry 250:7359–7365
    [Google Scholar]
  17. Nakae T., Ishii J., Tokunaga M. 1979; Subunit structure of functional porin oligomers that form permeability channels in the outer membrane of Escherichia coli. Journal of Biological Chemistry 254:1457–1461
    [Google Scholar]
  18. Nakamura K., Mizushima S. 1976; Effects of heating in dodecyl sulphate solution on the confor-mation and electrophoretic mobility of isolated major outer membrane proteins from Escherichia coli K-12. Journal of Biochemistry 80:1411–1422
    [Google Scholar]
  19. Ouchterlony Ö. 1958; Diffusion-in-gel methods for immunological analysis. In Progress in Allergy 5 pp. 1–78 Edited by Kallos P. Basel: S. Karger;
    [Google Scholar]
  20. Overbeeke N., Van Scharrenburg G., Lugtenberg B. 1980; Antigenic relationship between pore proteins of Escherichia coli K12. European Journal of Biochemistry 110:247–254
    [Google Scholar]
  21. Palva E. T., Randall L. L. 1978; Arrangement of protein I in Escherichia coli outer membrane: cross-linking study. Journal of Bacteriology 133:279–286
    [Google Scholar]
  22. Rosenbusch J. P. 1974; Characterization of the major envelope protein from Escherichia coli.Regular arrangement on the peptidoglycan and unusual dodecyl sulfate binding. Journal of Biological Chemistry 249:8019–8029
    [Google Scholar]
  23. Schnaitman C. A. 1971; Solubilization of the cytoplasmic membrane of Escherichia coli by Triton X-100. Journal of Bacteriology 108:545–552
    [Google Scholar]
  24. Tokunaga M., Tokunaga H., Okajima Y., Nakae T. 1979; Characterization of porins from the outer membrane of Salmonella typhimurium. 2. Physical properties of the functional oligomeric aggregates. European Journal of Biochemistry 95:441–448
    [Google Scholar]
  25. Van Tol M. J. D., Hofstra H., Dankert J. 1979; Major outer membrane proteins of Escherichia colianalysed by crossed immunoelectrophoresis. FEMS Microbiology Letters 5:349–352
    [Google Scholar]
  26. Verhoef C., De Graaff P. J., Lugtenberg E. J. J. 1977; Mapping of a gene for a major outer membrane protein of Escherichia coli K12 with the aid of a newly isolated bacteriophage. Molecular and General Genetics 150:103–105
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-125-2-285
Loading
/content/journal/micro/10.1099/00221287-125-2-285
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error