Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1976 Jun;73(6):1941–1944. doi: 10.1073/pnas.73.6.1941

Homologous inhibitors from potato tubers of serine endopeptidases and metallocarboxypeptidases.

C M Hass, R Venkatakrishnan, C A Ryan
PMCID: PMC430423  PMID: 1064864

Abstract

A potent polypeptide inhibitor of chymotrypsin has been purified from Russett Burbank potatoes. The inhibitor has no effect on bovine carboxypeptidases A or B but exhibits homology with a carboxypeptidase inhibitor that is also present in potato tubers. The chymotrypsin inhibitor has a molecular weight of approximately 5400 as estimated by gel filtration, amino acid analysis, and titration with chymotrypsin. The polypeptide chain consists of 49 amino acid residues, of which six are half-cystine, forming three disulfide bonds. Its size is similar to that of the carboxypeptidase inhibitor, which contains 39 amino acid residues and also has three disulfide bridges. In immunological double diffusion assays, the chymotrypsin inhibitor and the carboxypeptidase inhibitor do not crossreact; however, automatic Edman degradation of reduced and alkylated derivatives of the chymotrypsin inhibitor, yielding a partial sequence of 18 amino acid residues at the NH2-terminus, reveals a similarity in sequence to that of the carboxypeptidase inhibitor. Thus, inhibitors directed toward two distinct classes of proteases, the serine endopeptidases and the metallocarboxypeptidases, appear to have evolved from a common ancestor.

Full text

PDF
1941

Images in this article

1942Image
on p.1942

Selected References

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

  1. Axén R., Porath J., Ernback S. Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides. Nature. 1967 Jun 24;214(5095):1302–1304. doi: 10.1038/2141302a0. [DOI] [PubMed] [Google Scholar]
  2. CRESTFIELD A. M., MOORE S., STEIN W. H. The preparation and enzymatic hydrolysis of reduced and S-carboxymethylated proteins. J Biol Chem. 1963 Feb;238:622–627. [PubMed] [Google Scholar]
  3. Cavins J. F., Friedman M. An internal standard for amino acid analyses: S-beta-(4-pyridylethyl)-L-cysteine. Anal Biochem. 1970 Jun;35(2):489–493. doi: 10.1016/0003-2697(70)90211-3. [DOI] [PubMed] [Google Scholar]
  4. Creighton T. E. Homology of protein structures: proteinase inhibitors. Nature. 1975 Jun 26;255(5511):743–745. doi: 10.1038/255743a0. [DOI] [PubMed] [Google Scholar]
  5. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  6. ELLMAN G. L. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959 May;82(1):70–77. doi: 10.1016/0003-9861(59)90090-6. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Edman P., Begg G. A protein sequenator. Eur J Biochem. 1967 Mar;1(1):80–91. doi: 10.1007/978-3-662-25813-2_14. [DOI] [PubMed] [Google Scholar]
  9. FOLK J. E., SCHIRMER E. W. THE PORCINE PANCREATIC CARBOXYPEPTIDASE A SYSTEM. I. THREE FORMS OF THE ACTIVE ENZYME. J Biol Chem. 1963 Dec;238:3884–3894. [PubMed] [Google Scholar]
  10. HUMMEL B. C. A modified spectrophotometric determination of chymotrypsin, trypsin, and thrombin. Can J Biochem Physiol. 1959 Dec;37:1393–1399. [PubMed] [Google Scholar]
  11. Hass G. M., Nau H., Biemann K., Grahn D. T., Ericsson L. H., Neurath H. The amino acid sequence of a carboxypeptidase inhibitor from potatoes. Biochemistry. 1975 Mar 25;14(6):1334–1342. doi: 10.1021/bi00677a036. [DOI] [PubMed] [Google Scholar]
  12. Hermodson M. A., Ericsson L. H., Titani K., Neurath H., Walsh K. A. Application of sequenator analyses to the study of proteins. Biochemistry. 1972 Nov 21;11(24):4493–4502. doi: 10.1021/bi00774a011. [DOI] [PubMed] [Google Scholar]
  13. Huber R., Bode W., Kukla D., Kohl U., Ryan C. A. The structure of the complex formed by bovine trypsin and bovine pancreatic trypsin inhibitor III. Structure of the anhydro-trypsin-inhibitor complex. Biophys Struct Mech. 1975 May 30;1(3):189–201. doi: 10.1007/BF00535756. [DOI] [PubMed] [Google Scholar]
  14. Huber R., Kukla D., Bode W., Schwager P., Bartels K., Deisenhofer J., Steigemann W. Structure of the complex formed by bovine trypsin and bovine pancreatic trypsin inhibitor. II. Crystallographic refinement at 1.9 A resolution. J Mol Biol. 1974 Oct 15;89(1):73–101. doi: 10.1016/0022-2836(74)90163-6. [DOI] [PubMed] [Google Scholar]
  15. Melville J. C., Ryan C. A. Chymotrypsin inhibitor I from potatoes. Large scale preparation and characterization of its subunit components. J Biol Chem. 1972 Jun 10;247(11):3445–3453. [PubMed] [Google Scholar]
  16. Rancour J. M., Ryan C. A. Isolation of a carboxypeptidase B inhibitor from potattoes. Arch Biochem Biophys. 1968 Apr;125(1):380–383. doi: 10.1016/0003-9861(68)90675-9. [DOI] [PubMed] [Google Scholar]
  17. Reddy M. N., Keim P. S., Heinrikson R. L., Kezdy F. J. Primary structural analysis of sulfhydryl protease inhibitors from pineapple stem. J Biol Chem. 1975 Mar 10;250(5):1741–1750. [PubMed] [Google Scholar]
  18. Ryan C. A., Hass G. M., Kuhn R. W. Purification and properties of a carboxypeptidase inhibitor from potatoes. J Biol Chem. 1974 Sep 10;249(17):5495–5499. [PubMed] [Google Scholar]
  19. Sankoff D. Matching sequences under deletion-insertion constraints. Proc Natl Acad Sci U S A. 1972 Jan;69(1):4–6. doi: 10.1073/pnas.69.1.4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Smillie L. B., Enenkel A. G., Kay C. M. Physicochemical properties and amino acid composition of chymotrypsinogen B. J Biol Chem. 1966 May 10;241(9):2097–2102. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES