1887

Abstract

The major outer-membrane proteins of 40-41 kDa were identified by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) in strains ATCC 10953, ATCC 25586, F3, F6 and Fevl. The proteins were purified by preparative gel electrophoresis. Their behaviour in gel filtration and gel electrophoresis, their sensitivity to proteolytic enzymes, and their amino acid composition were investigated. The purified proteins were partly sequenced from the N-terminal end. A 36·5 kDa portion was protected against extrinsic proteolytic (trypsin, chymotrypsin or pronase) digestion of whole cells. This polypeptide was isolated and partially sequenced from the N-terminal end. From these data and data from extrinsic iodination it was concluded that the N-terminal end of the protein is probably exposed on the surface of the cell. A database search revealed amino acid sequence similarity in an Ala-Pro-rich region of outer-membrane protein A (OmpA) in other Gram-negative bacteria.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-135-12-3253
1989-12-01
2024-04-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/135/12/mic-135-12-3253.html?itemId=/content/journal/micro/10.1099/00221287-135-12-3253&mimeType=html&fmt=ahah

References

  1. Ames G.F-L., Nikaido K. 1976; Two-dimensional gel electrophoresis of membrane proteins.. Biochemistry 15:616–623
    [Google Scholar]
  2. Bakken V., Jensen H. B. 1986; Outer membrane proteins of Fusobacterium nucleatum Fevl.. Journal oj General Microbiology 132:1069–1078
    [Google Scholar]
  3. Blake M. S., Gotschlich E. C. 1982; Purification and partial characterization of the major outer- membrane protein of Neisseria gonorrhoeae. . Infection and Immunity 36:277–283
    [Google Scholar]
  4. Buchanan T. M., Pearce W. A. 1979; Pathogenic aspects of outer membrane components of gramnegative bacteria.. In Bacterial Outer Membranes pp. 475–514 Inoye M. Edited by New York: John Wiley;
    [Google Scholar]
  5. Butler P.J.G., Harris J. I., Hartley B. S., Leberman R. 1969; The use of maleic anhydride for the reversible blocking of amino groups in polypeptide chains.. Biochemical Journal 112:679–689
    [Google Scholar]
  6. Capaldi R. A., Vanderkooi G. 1972; The low polarity of many membrane proteins.. Proceedings of the National Academy of Sciences of the United States of America 69:930–932
    [Google Scholar]
  7. Chen R., Schmidmayr W., Krämer C., Chen-Schmeisser U., Henning U. 1980; Primary structure of major outer membrane protein II (OmpA protein) of Escherichia coli K-l 2.. Proceedings of the National Academy of Sciences of the United States of America 77:4592–4596
    [Google Scholar]
  8. Dirienzo J. M., Rosan B. 1984; Isolation of a major cell envelope protein from Fusobacterium nucleatum. . Infection and Immunity 44:386–393
    [Google Scholar]
  9. Dirienzo J. M., Spieler E. L. 1983; Identification and characterization of the major cell envelope proteins of oral strains of Actinobacillus actinomy- cetemcomitans. . Infection and Immunity 39:253–261
    [Google Scholar]
  10. Dzink J. L., Socransky S. S., Haffajee A. D. 1988; The predominant cultivable microbiota of active and inactive lesions of destructive peridontal diseases.. Journal of Clinical Periodontology 15:316–323
    [Google Scholar]
  11. Gotschlich E. C., Seiff M., Blake M. S. 1987; The DNA sequence of the structural gene of gonococcal protein III and the flanking region containing a repetitive sequence.. Journal of Experimental Medicine 165:471–482
    [Google Scholar]
  12. Hatch F. T. 1965; Correlation of amino-acid composition with certain characteristics of proteins.. Nature, London 206:777–779
    [Google Scholar]
  13. Hewick R. M., Hunkapiller M. W., Hood L. E., Dreyer W. J. 1981; A gas-liquid solid phase peptide and protein sequenator.. Journal of Biological Chemistry 256:7990–7997
    [Google Scholar]
  14. Higgins R. C., Dahmus M. E. 1979; Rapid visualization of protein bands in preparative SDS- polyacrylamide gels.. Analytical Biochemistry 93:257–260
    [Google Scholar]
  15. Kaufman J., Dirienzo J. M. 1989; Isolation of a corncob (coaggregation) receptor polypeptide from Fusobacterium nucleatum. . Infection and Immunity 57:331–337
    [Google Scholar]
  16. Laemmli U. K. 1970; Cleavage of structural proteins during assembly of the head of bacteriophage T4.. Nature, London 227:680–685
    [Google Scholar]
  17. Lugtenberg B., Van Alphen L. 1983; Molecular architecture and functioning of the outer membrane of Escherichia coli and other gram-negative bacteria.. Biochimica et biophysica acta 737:51–115
    [Google Scholar]
  18. Marchalonis J. J. 1969; An enzymic method for trace iodination of immunoglobulins and other proteins.. Biochemical Journal 113:299–305
    [Google Scholar]
  19. Marshall T., Latner A. L. 1981; Incorporation of methylamine in an ultrasensitive silver stain for detecting protein in thick polyacrylamide gels.. Electrophoresis 2:228–235
    [Google Scholar]
  20. O’Farrell P. H. 1975; High resolution two-dimensional electrophoresis of proteins.. Journal of Biological Chemistry 250:4007–4021
    [Google Scholar]
  21. 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]
  22. Schnaitman C. A. 1971; Solubilization of the cytoplasmic membrane of Escherichia coli by Triton X-100.. Journal of Bacteriology 108:545–552
    [Google Scholar]
  23. Schweizer M., Hindennach I., Garten W., Henning U. 1978; Major proteins of the Escherichia coli outer cell envelope membrane. Interaction of protein II with lipopolysaccharide.. European Journal of Biochemistry 82:211–217
    [Google Scholar]
  24. Socransky S. S. 1970; Relationship of bacteria to the etiology of peridontal disease.. Journal of Dental Research 49:203–222
    [Google Scholar]
  25. Socransky S. S., Haffajee A. D., Dzink J. L. 1988; Relationship of subgingival microbial complexes to clinical features at the sampled sites.. Journal of Clinical Periodontology 15:440–444
    [Google Scholar]
  26. Takada H., Ogawa T., Yoshimura F., Otsuka K., Kokeguchi S., Kato K., Umemoto T., Kotani S. 1988; Immunobiological activities of a porin fraction isolated from Fusobacterium nucleatum ATCC 10953.. Infection and Immunity 56:855–863
    [Google Scholar]
  27. Towbin H„STAEHELIN, Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.. Proceedings of the National Academy of Sciences of the United States of America 76:4350–4354
    [Google Scholar]
  28. Tsai C. M., Frasch C. E. 1982; A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels.. Analytical Biochemistry 119:115–119
    [Google Scholar]
  29. Vasstrand E. N., Hofstad T., Endresen C., Jensen H. B. 1979; Demonstration of lanthonine as a natural constituent of the peptidoglycan of Fusobacterium nucleatum. . Infection and Immunity 25:775–780
    [Google Scholar]
  30. Vasstrand E., Jensen H. B., Miron T. 1980; Microbore single-column analysis of amino acids and amino sugars specific to bacterial cell wall peptidoglycans.. Analytical Biochemistry 105:154–158
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-135-12-3253
Loading
/content/journal/micro/10.1099/00221287-135-12-3253
Loading

Data & Media loading...

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