Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1975 Jun;122(3):1265–1273. doi: 10.1128/jb.122.3.1265-1273.1975

Three yeast proteins that specifically inhibit yeast proteases A, B, and C.

J F Lenney
PMCID: PMC246184  PMID: 238943

Abstract

Baker's yeast was found to contain inhibitors of yeast proteases A and C. These two proteins were partially purified, characterized, and compared with the previously described inhibitor of protease B. The A and B inhibitors were very thermostable and were extracted from intact yeast cells at 9k C. The A inhibitor appeared to be a protein with a molecular weight of about 22,000 which could be dissociated into two monomers or chains, both of which had a molecular weight of approximately 11,000. The protease C (carboxypeptidase Y)-inhibitor complex was purified and then partially disociated on an ion-exchange column. The free protease C inhibitor was very unstable, possibly because of destruction by a contaminating protease. Each inhibitor was specific for its corresponding protease and each inhibition was competitive. Whereas proteases A, B, and C destroyed the B inhibitor, only protease B had a pronounced destructive effect on the protease A inhibitor. Pepstatin was found to be a selective inhibitor of protease A, whereas chymostatin and antipain specifically inhibited protease B.

Full text

PDF
1265

Images in this article

1268Image
on p.1268

Selected References

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

  1. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Betz H., Hinze H., Holzer H. Isolation and properties of two inhibitors of proteinase B from yeast. J Biol Chem. 1974 Jul 25;249(14):4515–4521. [PubMed] [Google Scholar]
  3. Cabib E., Ulane R. Chitin synthetase activating factor from yeast, a protease. Biochem Biophys Res Commun. 1973 Jan 4;50(1):186–191. doi: 10.1016/0006-291x(73)91081-4. [DOI] [PubMed] [Google Scholar]
  4. Chen A. W., Miller J. J. Proteolytic activity of intact yeast cells during sporulation. Can J Microbiol. 1968 Sep;14(9):957–963. doi: 10.1139/m68-159. [DOI] [PubMed] [Google Scholar]
  5. Félix F., Brouillet N. Purification et propriétés de deux peptidases de levure de brasserie. Biochim Biophys Acta. 1966 Jul 6;122(1):127–144. [PubMed] [Google Scholar]
  6. Hasilik A., Holzer H. Participation of the tryptophan synthase inactivating system from yeast in the activation of chitin synthase. Biochem Biophys Res Commun. 1973 Jul 17;53(2):552–559. doi: 10.1016/0006-291x(73)90697-9. [DOI] [PubMed] [Google Scholar]
  7. Hayashi R., Aibara S., Hata T. A unique carboxypeptidase activity of yeast proteinase C. Biochim Biophys Acta. 1970 Aug 15;212(2):359–361. doi: 10.1016/0005-2744(70)90218-4. [DOI] [PubMed] [Google Scholar]
  8. Hayashi R., Moore S., Stein W. H. Carboxypeptidase from yeast. Large scale preparation and the application to COOH-terminal analysis of peptides and proteins. J Biol Chem. 1973 Apr 10;248(7):2296–2302. [PubMed] [Google Scholar]
  9. Holzer H., Katsunuma T., Schött E. G., Ferguson A. R., Hasilki A., Betz H. Studies on a tryptophan synthase inactivating system from yeast. Adv Enzyme Regul. 1973;11:53–60. doi: 10.1016/0065-2571(73)90008-3. [DOI] [PubMed] [Google Scholar]
  10. Juni E., Heym G. A. Properties of yeast pyruvate decarboxylase and their modification by proteolytic enzymes. II. Selective alteration by yeast proteases. Arch Biochem Biophys. 1968 Sep 20;127(1):89–100. doi: 10.1016/0003-9861(68)90205-1. [DOI] [PubMed] [Google Scholar]
  11. LENNEY J. F. A study of two yeast proteinases. J Biol Chem. 1956 Aug;221(2):919–930. [PubMed] [Google Scholar]
  12. 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]
  13. Lenney J. F., Dalbec J. M. Purification and properties of two proteinases from Saccharomyces cerevisiae. Arch Biochem Biophys. 1967 Apr;120(1):42–48. doi: 10.1016/0003-9861(67)90595-4. [DOI] [PubMed] [Google Scholar]
  14. Lenney J. F., Dalbec J. M. Yeast proteinase B: identification of the inactive form as an enzyme-inhibitor complex. Arch Biochem Biophys. 1969 Jan;129(1):407–409. doi: 10.1016/0003-9861(69)90194-5. [DOI] [PubMed] [Google Scholar]
  15. Lenney J. F., Matile P., Wiemken A., Schellenberg M., Meyer J. Activities and cellular localization of yeast proteases and their inhibitors. Biochem Biophys Res Commun. 1974 Oct 23;60(4):1378–1383. doi: 10.1016/0006-291x(74)90350-7. [DOI] [PubMed] [Google Scholar]
  16. Matern H., Hoffmann M., Holzer H. Isolation and characterization of the carboxypeptidase Y inhibitor from yeast. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4874–4878. doi: 10.1073/pnas.71.12.4874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Matile P., Wiemken A. The vacuole as the lysosome of the yeast cell. Arch Mikrobiol. 1967 Feb 20;56(2):148–155. doi: 10.1007/BF00408765. [DOI] [PubMed] [Google Scholar]
  18. Molano J., Gancedo C. Specific inactivation of fructose 1,6-bisphosphatase from Saccharomyces cerevisiae by a yeast protease. Eur J Biochem. 1974 May 2;44(1):213–217. doi: 10.1111/j.1432-1033.1974.tb03475.x. [DOI] [PubMed] [Google Scholar]
  19. Moore S. Amino acid analysis: aqueous dimethyl sulfoxide as solvent for the ninhydrin reaction. J Biol Chem. 1968 Dec 10;243(23):6281–6283. [PubMed] [Google Scholar]
  20. SCHLAMOWITZ M., PETERSON L. U. Studies on the optimum pH for the action of pepsin on "native" and denatured bovine serum albumin and bovine hemoglobin. J Biol Chem. 1959 Dec;234:3137–3145. [PubMed] [Google Scholar]
  21. Saheki T., Matsuda Y., Holzer H. Urification and characterization of macromolecular inhibitors of proteinase A from yeast. Eur J Biochem. 1974 Sep 1;47(2):325–332. doi: 10.1111/j.1432-1033.1974.tb03697.x. [DOI] [PubMed] [Google Scholar]
  22. Schött E. H., Holzer H. Purification and some properties of tryptophan synthase inactivase II from yeast. Eur J Biochem. 1974 Feb 15;42(1):61–66. doi: 10.1111/j.1432-1033.1974.tb03314.x. [DOI] [PubMed] [Google Scholar]
  23. Tsai H., Tsai J. H., Yu P. H. Effects of yeast proteinase and its inhibitor on the inactivation of tryptophan synthase from Saccharomyces cerevisiae and Neurospora crassa. Eur J Biochem. 1973 Dec 3;40(1):225–232. doi: 10.1111/j.1432-1033.1973.tb03190.x. [DOI] [PubMed] [Google Scholar]
  24. Weber K., Pringle J. R., Osborn M. Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods Enzymol. 1972;26:3–27. doi: 10.1016/s0076-6879(72)26003-7. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES