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. 1981 Sep;147(3):1077–1084. doi: 10.1128/jb.147.3.1077-1084.1981

Purification and disposition of a surface protein associated with virulence of Aeromonas salmonicida.

W W Kay, J T Buckley, E E Ishiguro, B M Phipps, J P Monette, T J Trust
PMCID: PMC216148  PMID: 7275932

Abstract

Virulent strains of Aeromonas salmonicida observed by electron microscopy were characterized by an outer layer exhibiting a tetragonal repeat pattern. Attenuated strains had a 2.5 X 10(3)- to 5 X 10(3)-fold reduction in virulence and lost the outer layer, autoaggregating properties, and a 49-kilodalton protein (A protein) simultaneously. The A protein is the major protein component of outer membrane fractions of virulent strains. A variety of radiolabeling studies showed that this protein was surface localized and that it provided an effective barrier against iodination of other outer membrane proteins with either lactoperoxidase or diazoiodosulfanilic acid; A protein was not labeled with lactoperoxidase but was specifically labeled with diazoidosulfanilic acid. The A protein was purified by selective extraction with detergent and guanidine hydrochloride, and its amino acid composition was determined. The properties of A protein are compared with those of other bacterial surface layer proteins.

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Selected References

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

  1. Berg H. C. Sulfanilic acid diazonium salt: a label for the outside of the human erythrocyte membrane. Biochim Biophys Acta. 1969 Jun 3;183(1):65–78. doi: 10.1016/0005-2736(69)90130-8. [DOI] [PubMed] [Google Scholar]
  2. Buckmire F. L., Murray R. G. Studies on the cell wall of Spirillum serpens. 1. Isolation and partial purification of the outermost cell wall layer. Can J Microbiol. 1970 Oct;16(10):1011–1022. doi: 10.1139/m70-171. [DOI] [PubMed] [Google Scholar]
  3. Buckmire F. L., Murray R. G. Studies on the cell wall of Spirillum serpens. II. Chemical characterization of the outer structured layer. Can J Microbiol. 1973 Jan;19(1):59–66. doi: 10.1139/m73-009. [DOI] [PubMed] [Google Scholar]
  4. Every D., Skerman T. M. Ultrastructure of the Bacteroides nodosus cell envelope layers and surface. J Bacteriol. 1980 Feb;141(2):845–857. doi: 10.1128/jb.141.2.845-857.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  6. Filip C., Fletcher G., Wulff J. L., Earhart C. F. Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol. 1973 Sep;115(3):717–722. doi: 10.1128/jb.115.3.717-722.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. HOWARD A. N., WILD F. The reactions of diazonium compounds with amino acids and proteins. Biochem J. 1957 Apr;65(4):651–659. doi: 10.1042/bj0650651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hancock R. E., Carey A. M. Outer membrane of Pseudomonas aeruginosa: heat- 2-mercaptoethanol-modifiable proteins. J Bacteriol. 1979 Dec;140(3):902–910. doi: 10.1128/jb.140.3.902-910.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hastie A. T., Brinton C. C., Jr Isolation, characterization, and in vitro assembly of the tetragonally arrayed layer of Bacillus sphaericus. J Bacteriol. 1979 Jun;138(3):999–1009. doi: 10.1128/jb.138.3.999-1009.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hastie A. T., Brinton C. C., Jr Specific interaction of the tetragonally arrayed protein layer of Bacillus sphaericus with its peptidoglycan sacculus. J Bacteriol. 1979 Jun;138(3):1010–1021. doi: 10.1128/jb.138.3.1010-1021.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Marchalonis J. J., Cone R. E., Santer V. Enzymic iodination. A probe for accessible surface proteins of normal and neoplastic lymphocytes. Biochem J. 1971 Oct;124(5):921–927. doi: 10.1042/bj1240921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Markwell M. A., Fox C. F. Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril. Biochemistry. 1978 Oct 31;17(22):4807–4817. doi: 10.1021/bi00615a031. [DOI] [PubMed] [Google Scholar]
  13. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  14. McCoy E. C., Doyle D., Burda K., Corbeil L. B., Winter A. J. Superficial antigens of Campylobacter (Vibrio) fetus: characterization of antiphagocytic component. Infect Immun. 1975 Mar;11(3):517–525. doi: 10.1128/iai.11.3.517-525.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Neville D. M., Jr Molecular weight determination of protein-dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer system. J Biol Chem. 1971 Oct 25;246(20):6328–6334. [PubMed] [Google Scholar]
  16. Reyn A., Birch-Andersen A., Murray R. G. The fine structure of Cardiobacterium hominis. Acta Pathol Microbiol Scand B Microbiol Immunol. 1971;79(1):51–60. doi: 10.1111/j.1699-0463.1971.tb00032.x. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Sleytr U. B., Thorne K. J. Chemical characterization of the regularly arranged surface layers of Clostridium thermosaccharolyticum and Clostridium thermohydrosulfuricum. J Bacteriol. 1976 Apr;126(1):377–383. doi: 10.1128/jb.126.1.377-383.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Stewart M., Beveridge T. J., Murray R. G. Structure of the regular surface layer of Spirillum putridiconchylium. J Mol Biol. 1980 Feb 15;137(1):1–8. doi: 10.1016/0022-2836(80)90153-9. [DOI] [PubMed] [Google Scholar]
  20. Stewart M., Beveridge T. J. Structure of the regular surface layer of Sporosarcina ureae. J Bacteriol. 1980 Apr;142(1):302–309. doi: 10.1128/jb.142.1.302-309.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Swanson J., King G., Zeligs B. Studies on gonococcus infection. VIII. 125Iodine labeling of gonococci and studies on their in vitro interactions with eukaryotic cells. Infect Immun. 1975 Mar;11(3):453–459. doi: 10.1128/iai.11.3.453-459.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Thornley M. J., Thorne K. J., Glauert A. M. Detachment and chemical characterization of the regularly arranged subunits from the surface of an Acinetobacter. J Bacteriol. 1974 May;118(2):654–662. doi: 10.1128/jb.118.2.654-662.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Winter A. J., McCoy E. C., Fullmer C. S., Burda K., Bier P. J. Microcapsule of Campylobacter fetus: chemical and physical characterization. Infect Immun. 1978 Dec;22(3):963–971. doi: 10.1128/iai.22.3.963-971.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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