1887

Microbiology Society logo

Volume 135, Issue 12

Antigenic and Structural Analysis of Free

Abstract

Polypeptide and Western immunoblot profiles of subcellular fractions of ATCC 33520 have been determine by SDS-PAGE of Triton X-100-soluble and -insoluble fractions, a lipopolysaccharide-enriched fraction and purified flagella. Major Triton X-100-soluble polypeptides of 72, 68, 54 and 52 kDa were detected. The 54 kDa polypeptide appeared to be a breakdown product o a larger, heat-modifiable polypeptide. Based on the results o SDS-PAGE analysis and immunoblotting of proteinase K digests of , a ‘rough’ lipopolysaccharide appeared to be present. Electron microscopy has been used to monitor the effect of detergent treatment on the morphology o the organism and to examine the detailed structure of the flagella. Treatment with Triton removed the outer membrane, resulting in exposure of the flagella. The flagella were shown to have a complex sheath an core structure and polypeptide composition characteristic o that observed for other treponemes. Polypeptides of 38, 35, 32 and 28 kDa were present in purified flagella preparations. Immunoelectron microscopy, iodine-labelling and Western blotting were used to demonstrate the exposure o antigens on the surface. Surface iodination located polypeptides of 72, 68 and 54 kDa. Antiserum raised against whole cells of recognized these polypeptides and an additional polypeptide of 52 kDa. These data provide a basis for future detaile molecular analysis of the ultrastructure an antigenicity of .

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
© Society for General Microbiology 1989
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-135-12-3209
1989-12-01
2023-06-02
Loading full text...

Full text loading...

/deliver/fulltext/micro/135/12/mic-135-12-3209.html?itemId=/content/journal/micro/10.1099/00221287-135-12-3209&mimeType=html&fmt=ahah

References

  1. Armitage G. C., Dickinson W. R., Jenderseck R. S., Levine S. M., Chambers D. W. 1982; Relationship between the percentage of subgingival spirochetes and the severity of periodontal disease.. Journal of Periodontology 53:550–556
     [Google Scholar]
  2. Bailey M. J., Penn C. W., Cockayne A. 1985; Evidence for the presence of lipopolysaccharide in Treponema phagedenis (biotype Reiterii) but not in Treponema pallidum (Nichols).. FEMS Microbiology Letters 27:117–121
     [Google Scholar]
  3. Bailey M. J., Cockayne A., Penn C. W. 1987a; Monoclonal antibodies directed against surface-associated polypeptides of Treponema pallidum define a biologically active antigen.. Journal of General Microbiology 133:1793–1803
     [Google Scholar]
  4. Bretz W. A., Loesche W. J. 1987; Characteristics of trypsin-like activity in subgingival plaque samples.. Journal of Dental Research 66:1668–1672
     [Google Scholar]
  5. Cockayne A., Bailey M. J., Penn C. W. 1987; Analysis of sheath and core structures of the axial filament of Treponema pallidum.. Journal of General Microbiology 133:1397–1407
     [Google Scholar]
  6. Cheng S. L., Siboo R., Chinquee T., Johnson J. L., Mayberry W. R., Chan E.C.S. 1985; Comparative study of six random oral spirochete isolates: serological heterogeneity of Treponema denticola.. Journal of Periodontal Research 20:602–612
     [Google Scholar]
  7. Fehniger T. T., Walfield A. M., Cunningham T. M., Radolf J. D., Miller J. N., Lovett M. A. 1984; Purification and characterisation of a cloned protease-resistant Treponema pallidum-specific antigen.. Infection and Immunity 46:598–607
     [Google Scholar]
  8. Fukumoto Y., Okuda K., Kato T., Ohta K., Takazoe I. 1987; Taxonomic study of spirochetes isolated from human periodontal lesions.. Oral Microbiology and Immunology 2:82–87
     [Google Scholar]
  9. Geis G., Leying H., Suerbaum S., Mai U., Opferkuch W. 1989; Ultrastructure and chemical analysis of Camplyobacter pylori flagella.. Journal of Clinical Microbiology 27:436–441
     [Google Scholar]
  10. Hancock R.E.W. 1987; Role of porins in outer membrane permeability.. Journal of Bacteriology 169:929–933
     [Google Scholar]
  11. Hitchcock P. J., Brown T. M. 1983; Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels.. Journal of Bacteriology 154:269–277
     [Google Scholar]
  12. Jacob E., Nauman R. K. 1982; Common antigens of Treponema denticola: chemical, physical, and serological characterisation.. Infection and Immunity 37:474–480
     [Google Scholar]
  13. Kropinski A. M., Parr T. R., Angus B. L., Hancock R.E.W., Ghiorse W. C., Greenberg E. P. 1987; Isolation of the outer membrane and characterisation of the major outer membrane protein from Spirochaeta aurantia.. Journal of Bacteriology 169:172–179
     [Google Scholar]
  14. Krupski G., Gotz R., Ober K., Pleier E., Schmitt R. 1985; Structure of complex flagellar filaments in Rhizobium meliloti.. Journal of Bacteriology 162:361–366
     [Google Scholar]
  15. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature, London 227:680–685
     [Google Scholar]
  16. Listgarten M. A., Levin S. 1981; Positive correlation between the proportions of subgingival spirochetes and motile bacteria and susceptibility of human subjects to periodontal deterioration.. Journal of Clinical Periodontology 8:122–138
     [Google Scholar]
  17. Loesche W. J., Laughon B. E. 1982; Role of spirochaetes in periodontal disease.. In Host-Parasite Interactions in Periodontal Disease, pp 2–15 Genco R. J., Mergenhagen S. E. Edited by Washington, DC: American Society for Microbiology;
     [Google Scholar]
  18. Loesche W. J., Syed S. A., Schmidt E., Morrison E. C. 1985; Bacterial profiles of subgingival plaques in periodontitis.. Journal of Periodontology 56:447–456
     [Google Scholar]
  19. Moore W.E.C., Holdeman L. V., Smibert R. M., Hash D. E., Burmeister J. A., Ranney R. R. 1982; Bacteriology of severe periodontitis in young human adults.. Infection and Immunity 38:1137–1148
     [Google Scholar]
  20. Norris S. J., Charon N. W., Cook R. J., Fuentes M. D., Limberger R. J. 1988; Antigenic relatedness and N-terminal sequence homology define two classes of periplasmic flagellar proteins of Treponema pallidum subsp. pallidum and Treponema phagedenis.. Journal of Bacteriology 170:4072–4082
     [Google Scholar]
  21. Penn C. W., Bailey M. J., Cockayne A. 1985a; The axial filament antigen of Treponema pallidum.. Immunology 54:635–641
     [Google Scholar]
  22. Penn C. W., Cockayne A., Bailey M. J. 1985b; The outer membrane of Treponema pallidum: biological significance and biochemical properties.. Journal of General Microbiology 131:2349–2357
     [Google Scholar]
  23. Radolf J. D., Blanco D. R., Miller J. N., Lovett M. A. 1986; Antigenic interrelationship between endoflagella of Treponema phagodenis biotype Reiter and Treponema pallidum: molecular characterisation of endoflagellar proteins.. Infection and Immunity 54:626–634
     [Google Scholar]
  24. Russell R.R.B., Peach S. L., Coleman G., Cohen B. 1983; Antibody responses to antigens of Streptococcus mutans in monkeys (Macaca fascicularis) immunized against dental caries.. Journal of General Microbiology 129:865–875
     [Google Scholar]
  25. Russell R.R.B., Coleman D., Dougan G. 1985; Expression of a gene for glucan-binding protein from Streptococcus mutans in Escherichia coli.. Journal of General Microbiology 131:295–299
     [Google Scholar]
  26. Sela M. N., Kornman K. S., Ebersole J. L., Holt S. C. 1987; Characterisation of treponemes isolated from human and non human primate periodontal pockets.. Oral Microbiology and Immunology 2:21–29
     [Google Scholar]
  27. Simonson L. G., Goodman C. H., Bial J. J., Morton H. E. 1988a; Quantitative relationship of Treponema denticola to severity of periodontal disease.. Infection and Immunity 56:726–728
     [Google Scholar]
  28. Simonson L. G., Rouse R. F., Bockowski S. W. 1988b; Monoclonal antibodies that recognise a specific surface antigen of Treponema denticola.. Infection and Immunity 56:60–63
     [Google Scholar]
  29. Tall B. D., Nauman R. K. 1986; Microscopic agglutination and polyacrylamide gel electrophoresis analysis of oral anaerobic spirochetes.. Journal of Clinical Microbiology 24:282–287
     [Google Scholar]
  30. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.. Proceedings of the National Academy of Science of the United States of America 76:4350–4354
     [Google Scholar]
  31. Umemoto T., Zambon J. J., Genco R. J., Namikawa I. 1988; Major antigens of human oral spirochetes associated with periodontal disease.. Advances in Dental Research 2:292–296
     [Google Scholar]
  32. Umemoto T., Namikawa I., Suido H., Asai S. 1989; A major antigen of the outer envelope of a human oral spirochete - Treponema denticola.. Infection and Immunity 57:2470–2474
     [Google Scholar]
  33. Van Embden J.D.A., Van Der Donk R. V., Van Eijk H. G., Van Der Heide J. A., De Jong M. F., Van Olderen A. D., Osterhaus A. D., Schouls L. M. 1983; Molecular cloning and expression of Treponema pallidum DNA in Escherichia coli K-12.. Infection and Immunity 42:187–196
     [Google Scholar]
  34. Wray W., Boulikas T., Wray V. P., Hancock R. 1981; Silver staining of proteins in polyacrylamide gels.. Analytical Biochemistry 118:197–203
     [Google Scholar]
  35. Yang G.C.H., Schrank G. D., Freeman B. A. 1977; Purification of flagellar cores of Vibrio cholerae.. Journal of Bacteriology 129:1121–1128
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-135-12-3209
Loading
Antigenic and Structural Analysis of Treponema denticola
Microbiology 135, 3209 (1989); https://doi.org/10.1099/00221287-135-12-3209
/content/journal/micro/10.1099/00221287-135-12-3209
/content/journal/micro/10.1099/00221287-135-12-3209
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