1887

Abstract

SUMMARY:

Congo red was bound from solution by strains of Porphyromonas gingivalis including W50, HG189, HG184, NCTC 11834, Bg 381, WPH35, the slower brown pigmenting colonial variant W50/BR1, and the avirulent mutant W50/BE1, and by Porphyromonas endodontalis HG370 and Porphyromonas asaccharolytica B537. SDS-PAGE of whole cells of all species examined displayed a 66 kDa Congo-red-binding component which was also detected in the outer membranes of P. gingivalis W50 grown in the chemostat under both haemin limitation and haemin excess, and which corresponded to a Coomassie-blue-stained band of the same mobility. Pretreatment of haemin-excess batch-grown cells of W50 with polymyxin B, which binds to lipid A, did not inhibit binding, whilst binding was enhanced in the presence of 2 M ammonium sulphate, suggesting the involvement of non-specific hydrophobic interactions. Binding was also reduced by pretreatment with trypsin and papain, and by 8-anilino-1-naphthalenesulphonic acid, which binds to hydrophobic amino acids. The 66 kDa binding component was sensitive to proteinase K digestion, and loss of Congo red staining of this band correlated with the quantitative reduction in Congo red binding by whole cells. These data, and our previous work, show that Congo red and iron protoporphyrin IX (haemin) are bound to different outer-membrane components, and that Congo red binding may be of little value as a marker to detect virulent strains of P. gingivalis or those expressing haemin-binding proteins.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-141-1-205
1995-01-01
2024-05-06
Loading full text...

Full text loading...

/deliver/fulltext/micro/141/1/mic-141-1-205.html?itemId=/content/journal/micro/10.1099/00221287-141-1-205&mimeType=html&fmt=ahah

References

  1. Bramanti T. E., Holt S. C. 1990; Iron regulated outer membrane proteins in the periodontopathic bacterium. Bacteroides gingivalis. Biochem Biophys Res Commun 166:1146–1154
    [Google Scholar]
  2. Bramanti T. E., Holt S. C. 1991; Roles of porphyrins and host iron transport proteins in regulation of growth of Porphyromonas gingivalis . J Bacteriol 173:7330–7339
    [Google Scholar]
  3. Bramanti T. E., Holt S. C. 1992a; Effect of porphyrins and host iron transport proteins on outer membrane expression in Porphyromonas {Bacteroides) gingivalis : identification of a novel 26 kDa hemin-repressible surface protein . Microb Pathog 13:61–73
    [Google Scholar]
  4. Bramanti T. E., Holt S. C. 1992b; Localization of a Porphyromonas gingivalis 26-kilodalton heat-modifiable, hemin-regulated surface protein which translocates across the outer membrane . J Bacteriol 174:5827–5839
    [Google Scholar]
  5. Bramanti T. E., Holt S. C. 1993; Hemin uptake in Porphyromonas gingivalis : Omp26 is a hemin-binding surface protein . J Bacteriol 175:7413–7420
    [Google Scholar]
  6. Carman R., Ramakrishnan M. D., Harper F. H. 1990; Hemin levels in culture medium of Porphyromonas (Bacteroides) gingivalis regulate both hemin binding and trypsin-like protease production . Infect Immun 58:4016–4019
    [Google Scholar]
  7. Daskaleros P. A., Payne S. M. 1987; Congo red binding phenotype is associated with hemin binding and increased in-fectivity of Shigella flexneri in the HeLa cell model . Infect Immun 55:1393–1398
    [Google Scholar]
  8. Deneer H. G., Potter A. A. 1989; Effect of iron restriction on the outer membrane proteins of Actinobacillus (Haemophilus) pleuro-pneumoniae . Infect Immun 57:798–804
    [Google Scholar]
  9. Genco C. A., Odusanya B. M., Brown G. 1994; Binding and accumulation of hemin in Porphyromonas gingivalis are induced by hemin . Infect Immun 62:2885–2892
    [Google Scholar]
  10. Gibbons R. J., MacDonald J. B. 1960; Hemin and vitamin K compounds as required factors for the cultivation of certain strains of Bacteroides melaninogenicus . J Bacteriol 80:164–170
    [Google Scholar]
  11. Grenier D. 1991; Haemin-binding property of Porphyromonas gingivalis outer membranes . FEMS Microbiol ‘Lett 77:45–50
    [Google Scholar]
  12. Inoshita E., Iwakura K., Amano A., Tamagawa H., Shizukuishi S. 1991; Effect of transferrin on the growth of Porphyromonas gingivalis . J Dent Res 70:1258–1261
    [Google Scholar]
  13. Ishiguro E. E., Kay W. W., Ainsworth T., Chamberlain J. B., Austen R. A., Buckley J. T., Trust T. J. 1981; Loss of virulence during culture of Aeromonas salmonicida at high temperature . J Bacteriol 148:333–340
    [Google Scholar]
  14. Ishiguro E. E., Ainsworth T., Trust T. J., Kay W. W. 1985; Congo red, a differential medium for Aeromonas salmonicida , detects the presence of the cell surface protein array involved in virulence . J Bacteriol 164:1233–1237
    [Google Scholar]
  15. Jackson S., Burrows T. W. 1956; The pigmentation of Pasteurella pestis on a defined medium containing haemin . Br J Exp Path 37:577–583
    [Google Scholar]
  16. Kay W. W., Buckley J. T., Ishiguro E. E., Phipps B. M., Monette J. P. L., Trust T. J. 1981; Purification and disposition of a surface protein associated with virulence of Aeromonas salmonicida . J Bacteriol 147:1077–1084
    [Google Scholar]
  17. Kay W. W., Phipps B. M., Ishiguro E. E., Trust T. J. 1985; Porphyrin binding by the surface array virulence protein of Aeromonas salmonicida. J Bacteriol 164:1332–1336
    [Google Scholar]
  18. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  19. Mansheim B. J., Kasper D. L. 1977; Purification and immunological characterisation of the outer membrane complex of Bacteroides melaninogenicus subspecies asaccharolyticus . J Infect Dis 135:787–799
    [Google Scholar]
  20. Marsh P. D., McDermid A. S., McKee A. S., Baskerville A. 1994 The effect of growth rate and haemin on the virulence and proteolytic activity of Porphyromonas gingivalis W50 . Microbiology 140861–865
    [Google Scholar]
  21. Mayrand D., Bourgeau G., Grenier D., Lacroix J.-M. 1984; Properties of oral asaccharolytic black-pigmented Bacteroides . Can J Microbiol 30:1133–1136
    [Google Scholar]
  22. McKee A. S., McDermid A. S., Baskerville A., Dowsett A. B., Ell wood D. C., Marsh P. D. 1986; Effect of hemin on the physiology and virulence of Bacteroides gingivalis W50 . Infect Immun 52:349–355
    [Google Scholar]
  23. McKee A. S., McDermid A. S., Baskerville A., Marsh P. D. 1988; Isolation of colonial variants of Bacteroides gingivalis W50 with a reduced virulence . J Med Microbiol 27:59–64
    [Google Scholar]
  24. Minas T., Greenman J., Schaffer A. G. 1993; The influence of haemin levels on growth and enzyme production by Porphyromonas gingivalis in continuous culture . Microbios 76:105–114
    [Google Scholar]
  25. Papaioannou S., Marsh P. D., Ivanyi L. 1991; The immuno-genicity of outer membrane proteins of Porphyromonas (Bacteroides) gingivalis W50 in periodontal disease . Oral Microbiol Immunol 6:327–331
    [Google Scholar]
  26. Prpic J. K., Robins-Browne R. M., Davey R. B. 1983; Differentiation between virulent and avirulent Yersinia enterocolitica isolates using Congo red agar . J Clin Microbiol 18:486–490
    [Google Scholar]
  27. Scott D., Siboo I. R., Chan E. C. S., Klitorinas A., Siboo R. 1993; Binding of hemin and Congo red by oral hemolytic spirochetes. Oral Microbiol Immunol 8:245–250
    [Google Scholar]
  28. Smalley J. W., Birss A. J. 1987; Trypsin-like enzyme activity of the extracellular vesicles of Bacteroides gingivalis W50 . J Gen Microbiol 133:2883–2894
    [Google Scholar]
  29. Smalley J. W., Birss A. J., McKee A. S., Marsh P. D. 1991; Haemin-restriction influences haemin-binding, haemagglutination and protease activity of cells and extracellular membrane vesicles of Porphyromonas gingivalis W50 . FEMS Microbiol Lett 90:63–68
    [Google Scholar]
  30. Smalley J. W., Birss A. J., McKee A. S., Marsh P. D. 1993; Haemin-binding proteins of Porphyromonas gingivalis W50 grown in a chemostat under haemin-limitation . J Gen Microbiol 139:2145–2150
    [Google Scholar]
  31. Smalley J. W., Birss A. J., McKee A. S., Marsh P. D. 1994; Changes in the affinity of haemin-binding by Porphyromonas gingivalis W50 under different environmental conditions . Microb Ecol Hlth Dis 7:9–15
    [Google Scholar]
  32. Stugard C. E., Daskaleros P. A., Payne S. M. 1989; A 101-kilodalton heme-binding protein associated with Congo red binding and virulence of Shigella flexneri and enteroinvasive Escherichia coli strains . Infect Immun 7:3534–3539
    [Google Scholar]
  33. Surgalla M. J., Beesley E. D. 1969; Congo red agar plating medium for detecting pigmentation in Pasteurella pestis . . Appl Microbiol 18:834–837
    [Google Scholar]
  34. Wyss C. 1992; Growth of Porphyromonas gingivalis, Treponema denticola, T . pectinovorum, T. socranskii, and T. vincentii in a chemically defined medium . J Clin Microbiol 30:2225–2229
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-141-1-205
Loading
/content/journal/micro/10.1099/00221287-141-1-205
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