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. 1981 Sep;147(3):1095–1104. doi: 10.1128/jb.147.3.1095-1104.1981

Purification and characterization of surface fibrils from taxonomically typical Actinomyces viscosus WVU627.

N Masuda, R P Ellen, D A Grove
PMCID: PMC216150  PMID: 7275934

Abstract

Fibrils of Actinomyces viscosus WVU627 (numerical taxonomy cluster 1) were obtained by homogenization and purified by ultrafiltration, ammonium sulfate precipitations, gel filtration, and ion-exchange chromatography. Electron microscopy and resolution of a single band by sodium dodecyl sulfate-polyacrylamide gel electrophoresis attested to the purity of the preparation. Purified fibrils were composed mainly of protein; small quantities of carbohydrate and phosphorus were detected. Immunoelectrophoresis revealed only a single precipitable antigen, which migrated slightly toward the anode, in reactions between purified fibrils and antiserum raised against either whole bacterial cells or the purified fibrils themselves. Immunoelectron microscopy with ferritin-conjugated antifibril antibody hemagglutination inhibition, and bacterial agglutination tests demonstrated that fibrils of Actinomyces viscosus cluster 1 strains shared some common antigens with clusters 2, 3, 4 and 6, but did not cross-react with typical Actinomyces naeslundii of cluster 5. Stability tests revealed that after heat or alkali treatment, the fibrils lost their antigenicity and disappeared from electron micrographs. They were affected less by sodium dodecyl sulfate, sonic, or acid treatments.

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Selected References

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  1. Brinton C. C., Jr The structure, function, synthesis and genetic control of bacterial pili and a molecular model for DNA and RNA transport in gram negative bacteria. Trans N Y Acad Sci. 1965 Jun;27(8):1003–1054. doi: 10.1111/j.2164-0947.1965.tb02342.x. [DOI] [PubMed] [Google Scholar]
  2. Buchanan T. M. Antigenic heterogeneity of gonococcal pili. J Exp Med. 1975 Jun 1;141(6):1470–1475. doi: 10.1084/jem.141.6.1470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buchanan T. M., Swanson J., Holmes K. K., Kraus S. J., Gotschlich E. C. Quantitative determination of antibody to gonococcal pili. Changes in antibody levels with gonococcal infection. J Clin Invest. 1973 Nov;52(11):2896–2909. doi: 10.1172/JCI107486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cisar J. O., Kolenbrander P. E., McIntire F. C. Specificity of coaggregation reactions between human oral streptococci and strains of Actinomyces viscosus or Actinomyces naeslundii. Infect Immun. 1979 Jun;24(3):742–752. doi: 10.1128/iai.24.3.742-752.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cisar J. O., Vatter A. E. Surface fibrils (fimbriae) of Actinomyces viscosus T14V. Infect Immun. 1979 May;24(2):523–531. doi: 10.1128/iai.24.2.523-531.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clark W. B., Bammann L. L., Gibbons R. J. Comparative estimates of bacterial affinities and adsorption sites on hydroxyapatite surfaces. Infect Immun. 1978 Mar;19(3):846–853. doi: 10.1128/iai.19.3.846-853.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Costello A. H., Cisar J. O., Kolenbrander P. E., Gabriel O. Neuraminidase-dependent hamagglutination of human erythrocytes by human strains of Actinomyces viscosus and Actinomyces naeslundii. Infect Immun. 1979 Nov;26(2):563–572. doi: 10.1128/iai.26.2.563-572.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DISCHE Z., DEVI A. A new colorimetric method for the determination of ketohexoses in presence of aldoses, ketoheptoses and ketopentoses. Biochim Biophys Acta. 1960 Mar 25;39:140–144. doi: 10.1016/0006-3002(60)90129-3. [DOI] [PubMed] [Google Scholar]
  9. Ellen R. P., Balcerzak-Raczkowski I. B. Interbacterial aggregation of Actinomyces naeslundii and dental plaque streptococci. J Periodontal Res. 1977 Jan;12(1):11–20. doi: 10.1111/j.1600-0765.1977.tb00104.x. [DOI] [PubMed] [Google Scholar]
  10. Ellen R. P. Establishment and distribution of Actinomyces viscosus and Actinomyces naeslundii in the human oral cavity. Infect Immun. 1976 Nov;14(5):1119–1124. doi: 10.1128/iai.14.5.1119-1124.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ellen R. P., Fillery E. D., Chan K. H., Grove D. A. Sialidase-enhanced lectin-like mechanism for Actinomyces viscosus and Actinomyces naeslundii hemagglutination. Infect Immun. 1980 Feb;27(2):335–343. doi: 10.1128/iai.27.2.335-343.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ellen R. P., Walker D. L., Chan K. H. Association of long surface appendages with adherence-related functions of the gram-positive species Actinomyces naeslundii. J Bacteriol. 1978 Jun;134(3):1171–1175. doi: 10.1128/jb.134.3.1171-1175.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fillery E. D., Bowden G. H., Hardie J. M. A comparison of strains of bacteria designated Actinomyces viscosus and Actinomyces naeslundii. Caries Res. 1978;12(6):299–312. doi: 10.1159/000260349. [DOI] [PubMed] [Google Scholar]
  14. Girard A. E., Jacius B. H. Ultrastructure of Actinomyces viscosus and Actinomyces naeslundii. Arch Oral Biol. 1974 Jan;19(1):71–79. doi: 10.1016/0003-9969(74)90228-3. [DOI] [PubMed] [Google Scholar]
  15. Huis in 't Veld J. H., Linssen W. H. The localization of streptococcal group and type antigens: an electron-microscopic study using ferritin-labelled antisera. J Gen Microbiol. 1973 Feb;74(2):315–324. doi: 10.1099/00221287-74-2-315. [DOI] [PubMed] [Google Scholar]
  16. Kumazawa N., Yanagawa R. Chemical properties of the pili of Corynebacterium renale. Infect Immun. 1972 Jan;5(1):27–30. doi: 10.1128/iai.5.1.27-30.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kumazawa N., Yanagawa R. Comparison of the chemical and immunological properties of the pili of three types of Corynebacterium renale. Jpn J Microbiol. 1973 Jan;17(1):13–19. doi: 10.1111/j.1348-0421.1973.tb00698.x. [DOI] [PubMed] [Google Scholar]
  18. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  19. McIntire F. C., Vatter A. E., Baros J., Arnold J. Mechanism of coaggregation between Actinomyces viscosus T14V and Streptococcus sanguis 34. Infect Immun. 1978 Sep;21(3):978–988. doi: 10.1128/iai.21.3.978-988.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Reed M. J., Patters M. R., Mashimo P. A., Genco R. J., Levine R. J. Blastogenic response of human lymphocytes to oral bacterial antigens: characterization of bacterial sonicates. Infect Immun. 1976 Nov;14(5):1202–1212. doi: 10.1128/iai.14.5.1202-1212.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. SRI RAM J., TAWDE S. S., PIERCE G. B., Jr, MIDGLEY A. R., Jr Preparation of antibody-ferritin conjugates for immunoelectron microscopy. J Cell Biol. 1963 Jun;17:673–675. doi: 10.1083/jcb.17.3.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Salit I. E., Gotschlich E. C. Hemagglutination by purified type I Escherichia coli pili. J Exp Med. 1977 Nov 1;146(5):1169–1181. doi: 10.1084/jem.146.5.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Swanson J., Hsu K. C., Gotschlich E. C. Electron microscopic studies on streptococci. I. M antigen. J Exp Med. 1969 Nov 1;130(5):1063–1091. doi: 10.1084/jem.130.5.1063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  25. Wheeler T. T., Clark W. B. Fibril-mediated adherence of Actinomyces viscosus to saliva-treated hydroxyapatite. Infect Immun. 1980 May;28(2):577–584. doi: 10.1128/iai.28.2.577-584.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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