Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1995 Aug;63(8):3137–3142. doi: 10.1128/iai.63.8.3137-3142.1995

A conserved Aeromonas salmonicida porin provides protective immunity to rainbow trout.

P Lutwyche 1, M M Exner 1, R E Hancock 1, T J Trust 1
PMCID: PMC173428  PMID: 7622241

Abstract

A protein with an apparent M(r) of 28,000 was isolated from outer membrane preparations of Aeromonas salmonicida A440. The protein was tested for the ability to form pores, using a planar lipid bilayer model membrane system. The protein appeared to be a monomer with a single-channel conductance in 1.0 M KCl of 1.96 nS and a cation/anion permeability ratio of 2.91 +/- 0.68. These data show that the porin channel is comparable in size to OmpC and OmpF of Escherichia coli and is relatively nonselective, having some preference for cations over anions. The porin was purified by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and a polyclonal antibody was raised. Immunoblot analysis showed that an immunologically cross-reactive protein was present in other Aeromonas strains but not in strains of Vibrio or Yersinia. The N-terminal amino acid sequence of the porin was determined and was found to show some homology to an Aeromonas hydrophila outer membrane protein. This is the second porin species of A. salmonicida to be described, and it differs from the other in subunit molecular weight, aggregation properties, peptidoglycan association, pore size, and antigenicity. Rainbow trout (Oncorhynchus mykiss) immunized intraperitoneally with the purified porin protein were significantly protected from experimental A. salmonicida challenge. This is the first report of successful vaccination against A. salmonicida with a purified outer membrane component.

Full Text

The Full Text of this article is available as a PDF (407.8 KB).

Selected References

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

  1. Belland R. J., Trust T. J. Synthesis, export, and assembly of Aeromonas salmonicida A-layer analyzed by transposon mutagenesis. J Bacteriol. 1985 Sep;163(3):877–881. doi: 10.1128/jb.163.3.877-881.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benz R., Hancock R. E. Properties of the large ion-permeable pores formed from protein F of Pseudomonas aeruginosa in lipid bilayer membranes. Biochim Biophys Acta. 1981 Aug 20;646(2):298–308. doi: 10.1016/0005-2736(81)90336-9. [DOI] [PubMed] [Google Scholar]
  3. Benz R., Schmid A., Hancock R. E. Ion selectivity of gram-negative bacterial porins. J Bacteriol. 1985 May;162(2):722–727. doi: 10.1128/jb.162.2.722-727.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Darveau R. P., MacIntyre S., Buckley J. T., Hancock R. E. Purification and reconstitution in lipid bilayer membranes of an outer membrane, pore-forming protein of Aeromonas salmonicida. J Bacteriol. 1983 Dec;156(3):1006–1011. doi: 10.1128/jb.156.3.1006-1011.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DiRienzo J. M., Nakamura K., Inouye M. The outer membrane proteins of Gram-negative bacteria: biosynthesis, assembly, and functions. Annu Rev Biochem. 1978;47:481–532. doi: 10.1146/annurev.bi.47.070178.002405. [DOI] [PubMed] [Google Scholar]
  6. Doig P., Emödy L., Trust T. J. Binding of laminin and fibronectin by the trypsin-resistant major structural domain of the crystalline virulence surface array protein of Aeromonas salmonicida. J Biol Chem. 1992 Jan 5;267(1):43–49. [PubMed] [Google Scholar]
  7. Exner M. M., Doig P., Trust T. J., Hancock R. E. Isolation and characterization of a family of porin proteins from Helicobacter pylori. Infect Immun. 1995 Apr;63(4):1567–1572. doi: 10.1128/iai.63.4.1567-1572.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Garduño R. A., Kay W. W. Interaction of the fish pathogen Aeromonas salmonicida with rainbow trout macrophages. Infect Immun. 1992 Nov;60(11):4612–4620. doi: 10.1128/iai.60.11.4612-4620.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Granoff D. M., Munson R. S., Jr Prospects for prevention of Haemophilus influenzae type b disease by immunization. J Infect Dis. 1986 Mar;153(3):448–461. doi: 10.1093/infdis/153.3.448. [DOI] [PubMed] [Google Scholar]
  10. Heckels J. E., Fletcher J. N., Virji M. The potential protective effect of immunization with outer-membrane protein I from Neisseria gonorrhoeae. J Gen Microbiol. 1989 Aug;135(8):2269–2276. doi: 10.1099/00221287-135-8-2269. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Ishiguro E. E., Kay W. W., Ainsworth T., Chamberlain J. B., Austen R. A., Buckley J. T., Trust T. J. Loss of virulence during culture of Aeromonas salmonicida at high temperature. J Bacteriol. 1981 Oct;148(1):333–340. doi: 10.1128/jb.148.1.333-340.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jeanteur D., Gletsu N., Pattus F., Buckley J. T. Purification of Aeromonas hydrophila major outer-membrane proteins: N-terminal sequence analysis and channel-forming properties. Mol Microbiol. 1992 Nov;6(22):3355–3363. doi: 10.1111/j.1365-2958.1992.tb02203.x. [DOI] [PubMed] [Google Scholar]
  14. Kay W. W., Phipps B. M., Ishiguro E. E., Trust T. J. Porphyrin binding by the surface array virulence protein of Aeromonas salmonicida. J Bacteriol. 1985 Dec;164(3):1332–1336. doi: 10.1128/jb.164.3.1332-1336.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kennell W. L., Egli C., Hancock R. E., Holt S. C. Pore-forming ability of major outer membrane proteins from Wolinella recta ATCC 33238. Infect Immun. 1992 Feb;60(2):380–384. doi: 10.1128/iai.60.2.380-384.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Logan S. M., Trust T. J. Outer membrane characteristics of Campylobacter jejuni. Infect Immun. 1982 Dec;38(3):898–906. doi: 10.1128/iai.38.3.898-906.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marchesi V. T., Andrews E. P. Glycoproteins: isolation from cellmembranes with lithium diiodosalicylate. Science. 1971 Dec 17;174(4015):1247–1248. doi: 10.1126/science.174.4015.1247. [DOI] [PubMed] [Google Scholar]
  19. Michel C., Gonzalez R., Bonjour E., Avrameas S. A concurrent increasing of natural antibodies and enhancement of resistance to furunculosis in rainbow trout. Ann Rech Vet. 1990;21(3):211–218. [PubMed] [Google Scholar]
  20. Munn C. B., Ishiguro E. E., Kay W. W., Trust T. J. Role of surface components in serum resistance of virulent Aeromonas salmonicida. Infect Immun. 1982 Jun;36(3):1069–1075. doi: 10.1128/iai.36.3.1069-1075.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nikaido H. Porins and specific channels of bacterial outer membranes. Mol Microbiol. 1992 Feb;6(4):435–442. doi: 10.1111/j.1365-2958.1992.tb01487.x. [DOI] [PubMed] [Google Scholar]
  22. Phipps B. M., Kay W. W. Immunoglobulin binding by the regular surface array of Aeromonas salmonicida. J Biol Chem. 1988 Jul 5;263(19):9298–9303. [PubMed] [Google Scholar]
  23. Quinn D. M., Wong C. Y., Atkinson H. M., Flower R. L. Isolation of carbohydrate-reactive outer membrane proteins of Aeromonas hydrophila. Infect Immun. 1993 Feb;61(2):371–377. doi: 10.1128/iai.61.2.371-377.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Saint N., De E., Julien S., Orange N., Molle G. Ionophore properties of OmpA of Escherichia coli. Biochim Biophys Acta. 1993 Jan 18;1145(1):119–123. doi: 10.1016/0005-2736(93)90388-g. [DOI] [PubMed] [Google Scholar]
  26. Sugawara E., Nikaido H. Pore-forming activity of OmpA protein of Escherichia coli. J Biol Chem. 1992 Feb 5;267(4):2507–2511. [PubMed] [Google Scholar]
  27. Wilson M. E. The heat-modifiable outer membrane protein of Actinobacillus actinomycetemcomitans: relationship to OmpA proteins. Infect Immun. 1991 Jul;59(7):2505–2507. doi: 10.1128/iai.59.7.2505-2507.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Woodruff W. A., Parr T. R., Jr, Hancock R. E., Hanne L. F., Nicas T. I., Iglewski B. H. Expression in Escherichia coli and function of Pseudomonas aeruginosa outer membrane porin protein F. J Bacteriol. 1986 Aug;167(2):473–479. doi: 10.1128/jb.167.2.473-479.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES