Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1987 Mar;55(3):692–697. doi: 10.1128/iai.55.3.692-697.1987

Purification and biochemical properties of a bacteriocin from Actinobacillus actinomycetemcomitans.

R H Stevens, S E Lillard, B F Hammond
PMCID: PMC260395  PMID: 3818091

Abstract

Extracts of certain strains of Actinobacillus actinomycetemcomitans are inhibitory to strains of Streptococcus sanguis such as S. sanguis ATCC 10556. The isolation of a protein from an A. actinomycetemcomitans sonic extract which copurified with the inhibitory activity was accomplished by preparative isoelectric focusing, Sephadex G-100 gel filtration chromatography, and preparative polyacrylamide gel electrophoresis (PAGE). The resulting isolated protein, which focused at a pH of 6.1 to 6.3, appeared as a single band in anionic nondissociating PAGE analysis. This protein could be dissociated into two subunits with molecular weights of 50,000 and 70,000, which were resolvable by PAGE analysis. A 1,758-fold increase in specific activity was seen in the purified inhibitory protein compared with the crude sonic extract starting material. The properties of the inhibitory activity in the A. actinomycetemcomitans extract are characteristic of a bacteriocin. Accordingly, we propose the name actinobacillicin for the inhibitory protein.

Full text

PDF
692

Images in this article

694Image
on p.694
694Image
on p.694
695Image
on p.695
695Image
on p.695
696Image
on p.696

Selected References

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

  1. Beckers H. J., van der Hoeven J. S. Effect of microbial interaction on the colonization rate of Actinomyces viscosus or Streptococcus mutans in the dental plaque of rats. Infect Immun. 1982 Oct;38(1):8–13. doi: 10.1128/iai.38.1.8-13.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. De Graaf F. K., Klaasen-Boor P. Purification and characterization of a complex between cloacin and its immunity protein isolated from Enterobacter cloacae (Clo DF13). Dissociation and reconstitution of the complex. Eur J Biochem. 1977 Feb 15;73(1):107–114. doi: 10.1111/j.1432-1033.1977.tb11296.x. [DOI] [PubMed] [Google Scholar]
  3. Eisenmann A. C., Eisenmann R., Sousa O., Slots J. Microbiological study of localized juvenile periodontitis in Panama. J Periodontol. 1983 Dec;54(12):712–713. doi: 10.1902/jop.1983.54.12.712. [DOI] [PubMed] [Google Scholar]
  4. Hammond B. F., Lillard S. E., Stevens R. H. A bacteriocin of Actinobacillus actinomycetemcomitans. Infect Immun. 1987 Mar;55(3):686–691. doi: 10.1128/iai.55.3.686-691.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hardy K. G. Colicinogeny and related phenomena. Bacteriol Rev. 1975 Dec;39(4):464–515. doi: 10.1128/br.39.4.464-515.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hillman J. D., Johnson K. P., Yaphe B. I. Isolation of a Streptococcus mutans strain producing a novel bacteriocin. Infect Immun. 1984 Apr;44(1):141–144. doi: 10.1128/iai.44.1.141-144.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hu P. C., Brubaker R. R. Characterization of pesticin. Separation of antibacterial activities. J Biol Chem. 1974 Aug 10;249(15):4749–4753. [PubMed] [Google Scholar]
  8. Jakes K. S., Zinder N. D. Highly purified colicin E3 contains immunity protein. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3380–3384. doi: 10.1073/pnas.71.9.3380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jakes K., Zinder N. D., Boon T. Purification and properties of colicin E3 immunity protein. J Biol Chem. 1974 Jan 25;249(2):438–444. [PubMed] [Google Scholar]
  10. Jetten A. M., Vogels G. D., de Windt F. Production and purification of a Staphylococcus epidermidis bacteriocin. J Bacteriol. 1972 Oct;112(1):235–242. doi: 10.1128/jb.112.1.235-242.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Konisky J. Colicins and other bacteriocins with established modes of action. Annu Rev Microbiol. 1982;36:125–144. doi: 10.1146/annurev.mi.36.100182.001013. [DOI] [PubMed] [Google Scholar]
  12. Konisky J., Richards F. M. Characterization of colicin Ia and colicin Ib. Purification and some physical properties. J Biol Chem. 1970 Jun 10;245(11):2972–2978. [PubMed] [Google Scholar]
  13. Mandell R. L. A longitudinal microbiological investigation of Actinobacillus actinomycetemcomitans and Eikenella corrodens in juvenile periodontitis. Infect Immun. 1984 Sep;45(3):778–780. doi: 10.1128/iai.45.3.778-780.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mandell R. L., Socransky S. S. A selective medium for Actinobacillus actinomycetemcomitans and the incidence of the organism in juvenile periodontitis. J Periodontol. 1981 Oct;52(10):593–598. doi: 10.1902/jop.1981.52.10.593. [DOI] [PubMed] [Google Scholar]
  15. Moore W. E., Holdeman L. V., Cato E. P., Smibert R. M., Burmeister J. A., Palcanis K. G., Ranney R. R. Comparative bacteriology of juvenile periodontitis. Infect Immun. 1985 May;48(2):507–519. doi: 10.1128/iai.48.2.507-519.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moore W. E., Holdeman L. V., Smibert R. M., Good I. J., Burmeister J. A., Palcanis K. G., Ranney R. R. Bacteriology of experimental gingivitis in young adult humans. Infect Immun. 1982 Nov;38(2):651–667. doi: 10.1128/iai.38.2.651-667.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Moore W. E., Holdeman L. V., Smibert R. M., Hash D. E., Burmeister J. A., Ranney R. R. Bacteriology of severe periodontitis in young adult humans. Infect Immun. 1982 Dec;38(3):1137–1148. doi: 10.1128/iai.38.3.1137-1148.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Newman M. G., Socransky S. S. Predominant cultivable microbiota in periodontosis. J Periodontal Res. 1977 Mar;12(2):120–128. doi: 10.1111/j.1600-0765.1977.tb00114.x. [DOI] [PubMed] [Google Scholar]
  19. Newman M. G., Socransky S. S., Savitt E. D., Propas D. A., Crawford A. Studies of the microbiology of periodontosis. J Periodontol. 1976 Jul;47(7):373–379. doi: 10.1902/jop.1976.47.7.373. [DOI] [PubMed] [Google Scholar]
  20. Okuda K., Naito Y., Ohta K., Fukumoto Y., Kimura Y., Ishikawa I., Kinoshita S., Takazoe I. Bacteriological study of periodontal lesions in two sisters with juvenile periodontitis and their mother. Infect Immun. 1984 Jul;45(1):118–121. doi: 10.1128/iai.45.1.118-121.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pugsley A. P., Reeves P. Comparison of colicins B-K260 and D-CA23: purification and characterization of the colicins and examination of colicin immunity in the producing strains. Antimicrob Agents Chemother. 1977 Feb;11(2):345–358. doi: 10.1128/aac.11.2.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sano Y., Kageyama M. Purification and properties of an S-type pyocin, pyocin AP41. J Bacteriol. 1981 May;146(2):733–739. doi: 10.1128/jb.146.2.733-739.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schaller K., Nomura M. Colicin E2 is DNA endonuclease. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3989–3993. doi: 10.1073/pnas.73.11.3989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Slots J., Reynolds H. S., Genco R. J. Actinobacillus actinomycetemcomitans in human periodontal disease: a cross-sectional microbiological investigation. Infect Immun. 1980 Sep;29(3):1013–1020. doi: 10.1128/iai.29.3.1013-1020.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Slots J. Subgingival microflora and periodontal disease. J Clin Periodontol. 1979 Oct;6(5):351–382. doi: 10.1111/j.1600-051x.1979.tb01935.x. [DOI] [PubMed] [Google Scholar]
  26. Slots J. The predominant cultivable microflora of advanced periodontitis. Scand J Dent Res. 1977 Jan-Feb;85(2):114–121. doi: 10.1111/j.1600-0722.1977.tb00541.x. [DOI] [PubMed] [Google Scholar]
  27. Slots J. The predominant cultivable organisms in juvenile periodontitis. Scand J Dent Res. 1976 Jan;84(1):1–10. doi: 10.1111/j.1600-0722.1976.tb00454.x. [DOI] [PubMed] [Google Scholar]
  28. Slots J., Zambon J. J., Rosling B. G., Reynolds H. S., Christersson L. A., Genco R. J. Actinobacillus actinomycetemcomitans in human periodontal disease. Association, serology, leukotoxicity, and treatment. J Periodontal Res. 1982 Sep;17(5):447–448. doi: 10.1111/j.1600-0765.1982.tb02022.x. [DOI] [PubMed] [Google Scholar]
  29. Stevens R. H., Gatewood C., Hammond B. F. Cytotoxicity of the bacterium Actinobacillus actinomycetemcomitans extracts in human gingival fibroblasts. Arch Oral Biol. 1983;28(11):981–987. doi: 10.1016/0003-9969(83)90051-1. [DOI] [PubMed] [Google Scholar]
  30. Tagg J. R., Dajani A. S., Wannamaker L. W. Bacteriocins of gram-positive bacteria. Bacteriol Rev. 1976 Sep;40(3):722–756. doi: 10.1128/br.40.3.722-756.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Williams B. L., Ebersole J. L., Spektor M. D., Page R. C. Assessment of serum antibody patterns and analysis of subgingival microflora of members of a family with a high prevalence of early-onset periodontitis. Infect Immun. 1985 Sep;49(3):742–750. doi: 10.1128/iai.49.3.742-750.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Williams B. L., Pantalone R. M., Sherris J. C. Subgingival microflora and periodontitis. J Periodontal Res. 1976 Feb;11(1):1–18. doi: 10.1111/j.1600-0765.1976.tb00045.x. [DOI] [PubMed] [Google Scholar]
  33. Wolff L. F., Liljemark W. F., Bloomquist C. G., Pihlstrom B. L., Schaffer E. M., Bandt C. L. The distribution of Actinobacillus actinomycetemcomitans in human plaque. J Periodontal Res. 1985 May;20(3):237–250. doi: 10.1111/j.1600-0765.1985.tb00431.x. [DOI] [PubMed] [Google Scholar]
  34. Zambon J. J., Christersson L. A., Slots J. Actinobacillus actinomycetemcomitans in human periodontal disease. Prevalence in patient groups and distribution of biotypes and serotypes within families. J Periodontol. 1983 Dec;54(12):707–711. doi: 10.1902/jop.1983.54.12.707. [DOI] [PubMed] [Google Scholar]

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

RESOURCES