Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1983 Jan;39(1):311–314. doi: 10.1128/iai.39.1.311-314.1983

Effects of carrier ampholyte contamination on the biological and biochemical properties of streptococcal pyrogenic exotoxin type C.

T G Bloomster, D W Watson
PMCID: PMC347942  PMID: 6337098

Abstract

Three streptococcal pyrogenic exotoxins (SPEs), designated as SPE A, B, and C, have been purified and characterized. Routine purification of the SPEs includes the technique of isoelectric focusing. An earlier study showed that the removal of commercial carrier ampholytes (Ampholines) from SPE was difficult. The physiochemical properties of SPE C were previously reported; however, the SPE C preparation used in those experiments was contaminated with Ampholines. As an alternative to Ampholines, we used simple buffers to generate the isoelectric focusing pH gradient and used this SPE C in a comparative study to evaluate the effects of Ampholine contamination on the biological and biochemical properties of this toxin. We found that Ampholine contamination overestimates protein concentration; consequently, the biological activity of SPE C was actually greater than reported. The most serious effect of Ampholines in SPE C was on amino acid analysis. The presence of Ampholines causes an apparent increase in neutral amino acids and a decrease in basic amino acids.

Full text

PDF
311

Images in this article

313Image
on p.313

Selected References

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

  1. Barsumian E. L., Schlievert P. M., Watson D. W. Nonspecific and specific immunological mitogenicity by group A streptococcal pyrogenic exotoxins. Infect Immun. 1978 Dec;22(3):681–688. doi: 10.1128/iai.22.3.681-688.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bloomster T. G., Watson D. W. Detection of microgram quantities of carrier ampholytes in electrofocused proteins by thin-layer chromatography. Anal Biochem. 1981 May 1;113(1):79–84. doi: 10.1016/0003-2697(81)90046-4. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Cunningham C. M., Barsumian E. L., Watson D. W. Further purification of group A streptococcal pyrogenic exotoxin and characterization of the purified toxin. Infect Immun. 1976 Sep;14(3):767–775. doi: 10.1128/iai.14.3.767-775.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cunningham C. M., Watson D. W. Suppression of antibody response by group A streptococcal pyrogenic exotoxin and characterization of the cells involved. Infect Immun. 1978 Feb;19(2):470–476. doi: 10.1128/iai.19.2.470-476.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Edelhoch H. Spectroscopic determination of tryptophan and tyrosine in proteins. Biochemistry. 1967 Jul;6(7):1948–1954. doi: 10.1021/bi00859a010. [DOI] [PubMed] [Google Scholar]
  7. Hale M. L., Hanna E. E. Deregulation of mouse antibody-forming cells by streptococcal pyrogenic exotoxin. II. Modification of spleen T-cell-complemented nude mouse PFC responses. Cell Immunol. 1976 Oct;26(2):168–177. doi: 10.1016/0008-8749(76)90361-0. [DOI] [PubMed] [Google Scholar]
  8. Hanna E. E., Watson D. W. Enhanced immune response after immunosuppression by Streptococcal pyrogenic exotoxin. Infect Immun. 1973 Jun;7(6):1009–1011. doi: 10.1128/iai.7.6.1009-1011.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kim Y. B., Watson D. W. A purified group A streptococcal pyrogenic exotoxin. Physiochemical and biological properties including the enhancement of susceptibility to endotoxin lethal shock. J Exp Med. 1970 Mar 1;131(3):611–622. doi: 10.1084/jem.131.3.611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Prestidge R. L., Hearn M. T. Preparative flatbed electrofocusing in granulated gels with natural pH gradients generated from simple buffers. Anal Biochem. 1979 Aug;97(1):95–102. doi: 10.1016/0003-2697(79)90332-4. [DOI] [PubMed] [Google Scholar]
  11. Schlievert P. M., Bettin K. M., Watson D. W. Inhibition of ribonucleic acid synthesis by group A streptococcal pyrogenic exotoxin. Infect Immun. 1980 Feb;27(2):542–548. doi: 10.1128/iai.27.2.542-548.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Schlievert P. M., Bettin K. M., Watson D. W. Purification and characterization of group A streptococcal pyrogenic exotoxin type C. Infect Immun. 1977 May;16(2):673–679. doi: 10.1128/iai.16.2.673-679.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Schlievert P. M., Bettin K. M., Watson D. W. Reinterpretation of the Dick test: role of group A streptococcal pyrogenic exotoxin. Infect Immun. 1979 Nov;26(2):467–472. doi: 10.1128/iai.26.2.467-472.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Schlievert P. M., Schoettle D. J., Watson D. W. Nonspecific T-lymphocyte mitogenesis by pyrogenic exotoxins from group A streptococci and Staphylococcus aureus. Infect Immun. 1979 Sep;25(3):1075–1077. doi: 10.1128/iai.25.3.1075-1077.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schlievert P. M., Schoettle D. J., Watson D. W. Purification and physicochemical and biological characterization of a staphylococcal pyrogenic exotoxin. Infect Immun. 1979 Mar;23(3):609–617. doi: 10.1128/iai.23.3.609-617.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schlievert P. M., Watson D. W. Group A streptococcal pyrogenic exotoxin: pyrogenicity, alteration of blood-brain barrier, and separation of sites for pyrogenicity and enhancement of lethal endotoxin shock. Infect Immun. 1978 Sep;21(3):753–763. doi: 10.1128/iai.21.3.753-763.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. WATSON D. W. Host-parasite factors in group A streptococcal infections. Pyrogenic and other effects of immunologic distinct exotoxins related to scarlet fever toxins. J Exp Med. 1960 Feb 1;111:255–284. doi: 10.1084/jem.111.2.255. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES