Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1985 May 15;228(1):211–217. doi: 10.1042/bj2280211

Purification and characterization of a saliva-interacting cell-wall protein from Streptococcus mutans serotype f by using monoclonal-antibody immunoaffinity chromatography.

F Ackermans, J P Klein, J Ogier, H Bazin, F Cormont, R M Frank
PMCID: PMC1144971  PMID: 3890841

Abstract

A rat monoclonal antibody, LO SM2, of the immunoglobulin M class, specific for a saliva receptor (SR) from Streptococcus mutans serotype f, was able to precipitate the SR from crude cell-wall-associated antigens (WEA) of this bacteria in presence of a detergent mixture. We have then used the technique of monoclonal-antibody immunoaffinity chromatography to purify the S. mutans SR. Pure SR was obtained from a crude WEA fraction with a single chromatographic step. The active SR could be eluted from the column in a highly purified form with 0.2 M-glycine/HC1, pH 2.8. The final yield was about 32% in terms of binding activity. Characterization of the SR by crossed immunoelectrophoresis, sodium dodecyl sulphate- or 4-30%-native-gradient-polyacrylamide-gel electrophoresis showed that the receptor is a single polypeptide chain of Mr approx. 74000. Native or denaturated forms of the SR adsorbed on to a solid support, such as nitrocellulose, are recognized by monoclonal antibody LO SM2, and both forms are still able to bind the ligand, saliva.

Full text

PDF
211

Images in this article

214Fig. 1.
on p.214
215Fig. 2.
on p.215
216Fig. 3.
on p.216
217Fig. 4.
on p.217

Selected References

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

  1. Avrameas S., Guilbert B. A method for quantitative determination of cellular immunoglobulins by enzyme-labeled antibodies. Eur J Immunol. 1971 Nov;1(5):394–396. doi: 10.1002/eji.1830010518. [DOI] [PubMed] [Google Scholar]
  2. Bailyes E. M., Newby A. C., Siddle K., Luzio J. P. Solubilization and purification of rat liver 5'-nucleotidase by use of a zwitterionic detergent and a monoclonal-antibody immunoadsorbent. Biochem J. 1982 Apr 1;203(1):245–251. doi: 10.1042/bj2030245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bazin H., Cormont F., De Clercq L. Rat monoclonal antibodies. II. A rapid and efficient method of purification from ascitic fluid or serum. J Immunol Methods. 1984 Jun 8;71(1):9–16. doi: 10.1016/0022-1759(84)90200-x. [DOI] [PubMed] [Google Scholar]
  4. Cisar J. O., Barsumian E. L., Curl S. H., Vatter A. E., Sandberg A. L., Siraganian R. P. Detection and localization of a lectin on Actinomyces viscosus T14V by monoclonal antibodies. J Immunol. 1981 Oct;127(4):1318–1322. [PubMed] [Google Scholar]
  5. Clark W. B., Gibbons R. J. Influence of salivary components and extracellular polysaccharide synthesis from sucrose on the attachment of Streptococcus mutans 6715 to hydroxyapatite surfaces. Infect Immun. 1977 Nov;18(2):514–523. doi: 10.1128/iai.18.2.514-523.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Corthier G., Boschetti E., Charley-Poulain J. Improved method for IgG purification from various animal species by ion exchange chromatography. J Immunol Methods. 1984 Jan 20;66(1):75–79. doi: 10.1016/0022-1759(84)90249-7. [DOI] [PubMed] [Google Scholar]
  7. Denney R. M., Fritz R. R., Patel N. T., Abell C. W. Human liver MAO-A and MAO-B separated by immunoaffinity chromatography with MAO-B-specific monoclonal antibody. Science. 1982 Mar 12;215(4538):1400–1403. doi: 10.1126/science.7063850. [DOI] [PubMed] [Google Scholar]
  8. Ericson T., Rundegren J. Characterization of a salivary agglutinin reacting with a serotype c strain of Streptococcus mutans. Eur J Biochem. 1983 Jun 15;133(2):255–261. doi: 10.1111/j.1432-1033.1983.tb07456.x. [DOI] [PubMed] [Google Scholar]
  9. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  10. Gibbons R. J., Etherden I. Comparative hydrophobicities of oral bacteria and their adherence to salivary pellicles. Infect Immun. 1983 Sep;41(3):1190–1196. doi: 10.1128/iai.41.3.1190-1196.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gibbons R. J., Houte J. V. Bacterial adherence in oral microbial ecology. Annu Rev Microbiol. 1975;29:19–44. doi: 10.1146/annurev.mi.29.100175.000315. [DOI] [PubMed] [Google Scholar]
  12. Hamada S., Slade H. D. Biology, immunology, and cariogenicity of Streptococcus mutans. Microbiol Rev. 1980 Jun;44(2):331–384. doi: 10.1128/mr.44.2.331-384.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kuramitsu H., Ingersoll L. Molecular basis for the different sucrose-dependent adherence properties of Streptococcus mutans and Streptococcus sanguis. Infect Immun. 1977 Aug;17(2):330–337. doi: 10.1128/iai.17.2.330-337.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. McBride B. C., Gisslow M. T. Role of sialic acid in saliva-induced aggregation of Streptococcus sanguis. Infect Immun. 1977 Oct;18(1):35–40. doi: 10.1128/iai.18.1.35-40.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McBride B. C., Song M., Krasse B., Olsson J. Biochemical and immunological differences between hydrophobic and hydrophilic strains of Streptococcus mutans. Infect Immun. 1984 Apr;44(1):68–75. doi: 10.1128/iai.44.1.68-75.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ogier J. A., Klein J. P., Sommer P., Frank R. M. Identification and preliminary characterization of saliva-interacting surface antigens of Streptococcus mutans by immunoblotting, ligand blotting, and immunoprecipitation. Infect Immun. 1984 Jul;45(1):107–112. doi: 10.1128/iai.45.1.107-112.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Russell R. R. Wall-associated protein antigens of Streptococcus mutans. J Gen Microbiol. 1979 Sep;114(1):109–115. doi: 10.1099/00221287-114-1-109. [DOI] [PubMed] [Google Scholar]
  20. Schöller M., Klein J. P., Frank R. M. Common antigens of streptococcal and non-streptococcal oral bacteria: immunochemical studies of extracellular and cell-wall-associated antigens from Streptococcus sanguis, Streptococcus mutans, Lactobacillus salivarius, and Actinomyces viscosus. Infect Immun. 1981 Jan;31(1):52–60. doi: 10.1128/iai.31.1.52-60.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schöller M., Klein J. P., Sommer P., Frank R. Protoplast and cytoplasmic membrane preparations from Streptococcus sanguis and Streptococcus mutans. J Gen Microbiol. 1983 Oct;129(10):3271–3279. doi: 10.1099/00221287-129-10-3271. [DOI] [PubMed] [Google Scholar]
  22. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weeke B. A manual of quantitative immunoelectrophoresis. Methods and applications. 1. General remarks on principles, equipment, reagents and procedures. Scand J Immunol Suppl. 1973;1:15–35. doi: 10.1111/j.1365-3083.1973.tb03776.x. [DOI] [PubMed] [Google Scholar]
  24. Weerkamp A. H., Jacobs T. Cell wall-associated protein antigens of Streptococcus salivarius: purification, properties, and function in adherence. Infect Immun. 1982 Oct;38(1):233–242. doi: 10.1128/iai.38.1.233-242.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES