Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1989 Feb 15;258(1):305–308. doi: 10.1042/bj2580305

The inhibition of proinsulin-processing endopeptidase activities by active-site-directed peptides.

C J Rhodes 1, A Zumbrunn 1, E M Bailyes 1, E Shaw 1, J C Hutton 1
PMCID: PMC1138356  PMID: 2649090

Abstract

Inhibitor studies were performed on the two endopeptidase activities involved in proinsulin conversion in isolated insulin secretory granules [Davidson, Rhodes & Hutton (1988) Nature (London) 333, 93-96]. The active-site-directed peptides L-alanyl-L-arginyl-L-arginylmethyldimethylsulphonium and L-alanyl-L-lysyl-L-arginylmethyldimethylsulphonium inhibited these activities in accordance with the observed cleavage pattern, suggesting that the primary amino acid sequence of the dibasic site was an important determinant of the endopeptidase substrate specificities.

Full text

PDF
307

Selected References

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

  1. 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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Brennan S. O., Peach R. J. Calcium-dependent KEX2-like protease found in hepatic secretory vesicles converts proalbumin to albumin. FEBS Lett. 1988 Feb 29;229(1):167–170. doi: 10.1016/0014-5793(88)80819-6. [DOI] [PubMed] [Google Scholar]
  3. Chick W. L., Warren S., Chute R. N., Like A. A., Lauris V., Kitchen K. C. A transplantable insulinoma in the rat. Proc Natl Acad Sci U S A. 1977 Feb;74(2):628–632. doi: 10.1073/pnas.74.2.628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Davidson H. W., Hutton J. C. The insulin-secretory-granule carboxypeptidase H. Purification and demonstration of involvement in proinsulin processing. Biochem J. 1987 Jul 15;245(2):575–582. doi: 10.1042/bj2450575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davidson H. W., Peshavaria M., Hutton J. C. Proteolytic conversion of proinsulin into insulin. Identification of a Ca2+-dependent acidic endopeptidase in isolated insulin-secretory granules. Biochem J. 1987 Sep 1;246(2):279–286. doi: 10.1042/bj2460279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Davidson H. W., Rhodes C. J., Hutton J. C. Intraorganellar calcium and pH control proinsulin cleavage in the pancreatic beta cell via two distinct site-specific endopeptidases. Nature. 1988 May 5;333(6168):93–96. doi: 10.1038/333093a0. [DOI] [PubMed] [Google Scholar]
  7. Docherty K., Carroll R. J., Steiner D. F. Conversion of proinsulin to insulin: involvement of a 31,500 molecular weight thiol protease. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4613–4617. doi: 10.1073/pnas.79.15.4613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Docherty K., Carroll R., Steiner D. F. Identification of a 31,500 molecular weight islet cell protease as cathepsin B. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3245–3249. doi: 10.1073/pnas.80.11.3245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Docherty K., Hutton J. C. Carboxypeptidase activity in the insulin secretory granule. FEBS Lett. 1983 Oct 3;162(1):137–141. doi: 10.1016/0014-5793(83)81065-5. [DOI] [PubMed] [Google Scholar]
  10. Docherty K., Hutton J. C., Steiner D. F. Cathepsin B-related proteases in the insulin secretory granule. J Biol Chem. 1984 May 25;259(10):6041–6044. [PubMed] [Google Scholar]
  11. Docherty K., Steiner D. F. Post-translational proteolysis in polypeptide hormone biosynthesis. Annu Rev Physiol. 1982;44:625–638. doi: 10.1146/annurev.ph.44.030182.003205. [DOI] [PubMed] [Google Scholar]
  12. Hutton J. C., Davidson H. W., Peshavaria M. Proteolytic processing of chromogranin A in purified insulin granules. Formation of a 20 kDa N-terminal fragment (betagranin) by the concerted action of a Ca2+-dependent endopeptidase and carboxypeptidase H (EC 3.4.17.10). Biochem J. 1987 Jun 1;244(2):457–464. doi: 10.1042/bj2440457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hutton J. C., Penn E. J., Jackson P., Hales C. N. Isolation and characterization of calmodulin from an insulin-secreting tumour. Biochem J. 1981 Mar 1;193(3):875–885. doi: 10.1042/bj1930875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kemmler W., Steiner D. F. Conversion of proinsulin to insulin in a subcellular fraction from rat islets. Biochem Biophys Res Commun. 1970 Dec 9;41(5):1223–1230. doi: 10.1016/0006-291x(70)90217-2. [DOI] [PubMed] [Google Scholar]
  15. Loh Y. P., Brownstein M. J., Gainer H. Proteolysis in neuropeptide processing and other neural functions. Annu Rev Neurosci. 1984;7:189–222. doi: 10.1146/annurev.ne.07.030184.001201. [DOI] [PubMed] [Google Scholar]
  16. Moore H. P., Walker M. D., Lee F., Kelly R. B. Expressing a human proinsulin cDNA in a mouse ACTH-secreting cell. Intracellular storage, proteolytic processing, and secretion on stimulation. Cell. 1983 Dec;35(2 Pt 1):531–538. doi: 10.1016/0092-8674(83)90187-3. [DOI] [PubMed] [Google Scholar]
  17. Orci L. The insulin factory: a tour of the plant surroundings and a visit to the assembly line. The Minkowski lecture 1973 revisited. Diabetologia. 1985 Aug;28(8):528–546. doi: 10.1007/BF00281987. [DOI] [PubMed] [Google Scholar]
  18. Rauber P., Walker B., Stone S., Shaw E. Synthesis of lysine-containing sulphonium salts and their properties as proteinase inhibitors. Biochem J. 1988 Mar 15;250(3):871–876. doi: 10.1042/bj2500871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rhodes C. J., Lucas C. A., Mutkoski R. L., Orci L., Halban P. A. Stimulation by ATP of proinsulin to insulin conversion in isolated rat pancreatic islet secretory granules. Association with the ATP-dependent proton pump. J Biol Chem. 1987 Aug 5;262(22):10712–10717. [PubMed] [Google Scholar]
  20. Shaw E., Green G. D. Inactivation of thiol proteases with peptidyl diazomethyl ketones. Methods Enzymol. 1981;80(Pt 100):820–826. doi: 10.1016/s0076-6879(81)80064-x. [DOI] [PubMed] [Google Scholar]
  21. Shaw E. Peptidyl sulfonium salts. A new class of protease inhibitors. J Biol Chem. 1988 Feb 25;263(6):2768–2772. [PubMed] [Google Scholar]
  22. Sorenson R. L., Steffes M. W., Lindall A. W. Subcellular localization of proinsulin to insulin conversion in isolated rat islets. Endocrinology. 1970 Jan;86(1):88–96. doi: 10.1210/endo-86-1-88. [DOI] [PubMed] [Google Scholar]
  23. Thim L., Hansen M. T., Norris K., Hoegh I., Boel E., Forstrom J., Ammerer G., Fiil N. P. Secretion and processing of insulin precursors in yeast. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6766–6770. doi: 10.1073/pnas.83.18.6766. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES