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
. 1974 Apr;71(4):1070–1072. doi: 10.1073/pnas.71.4.1070

Studies on the Extended Active Sites of Acid Proteinases

P S Sampath-Kumar 1, J S Fruton 1,*
PMCID: PMC388164  PMID: 4598291

Abstract

The kinetics of the hydrolysis of a series of peptide substrates at a single peptide bond (between L-phenylalanyl and L-phenylalanyl) by the acid proteinases gastric pepsin A (EC 3.4.23.1), Rhizopus pepsin (EC 3.4.23.9), and beef-spleen cathepsin D (EC 3.4.23.5) have been determined by use of the fluorescamine assay method. The results indicate that the extended active site of pepsin can accommodate a sequence of at least seven amino-acid residues. Although the other two acid proteinases appear to act at the sensitive L-phenylalanyl-L-phenylalanyl bond by a mechanism similar to that of pepsin, the influence of structural changes on either side of the sensitive dipeptidyl unit on the kinetic parameters is different from that for pepsin. These data give further evidence for the importance of secondary interactions in determining the catalytic efficiency of enzymes that act on oligomeric substrates.

Keywords: pepsin, cathepsin D, Rhizopus pepsin, enzyme specificity, peptides

Full text

PDF
1070

Selected References

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

  1. Berger A., Schechter I. Mapping the active site of papain with the aid of peptide substrates and inhibitors. Philos Trans R Soc Lond B Biol Sci. 1970 Feb 12;257(813):249–264. doi: 10.1098/rstb.1970.0024. [DOI] [PubMed] [Google Scholar]
  2. Blake C. C., Johnson L. N., Mair G. A., North A. C., Phillips D. C., Sarma V. R. Crystallographic studies of the activity of hen egg-white lysozyme. Proc R Soc Lond B Biol Sci. 1967 Apr 18;167(1009):378–388. doi: 10.1098/rspb.1967.0035. [DOI] [PubMed] [Google Scholar]
  3. Ferguson J. B., Andrews J. R., Voynick I. M., Fruton J. S. The specificity of cathepsin D. J Biol Chem. 1973 Oct 10;248(19):6701–6708. [PubMed] [Google Scholar]
  4. Fruton J. S. The specificity and mechanism of pepsin action. Adv Enzymol Relat Areas Mol Biol. 1970;33:401–443. doi: 10.1002/9780470122785.ch9. [DOI] [PubMed] [Google Scholar]
  5. Graham J. E., Sodek J., Hofmann T. Rhizopus acid proteinases (rhizopus-pepsins): properties and homology with other acid proteinases. Can J Biochem. 1973 Jun;51(6):789–796. doi: 10.1139/o73-098. [DOI] [PubMed] [Google Scholar]
  6. Humphreys R. E., Fruton J. S. The substrate binding site of pepsin. Proc Natl Acad Sci U S A. 1968 Feb;59(2):519–525. doi: 10.1073/pnas.59.2.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Inouye K., Fruton J. S. The inhibition of pepsin action. Biochemistry. 1968 May;7(5):1611–1615. doi: 10.1021/bi00845a001. [DOI] [PubMed] [Google Scholar]
  8. Koshland D. E., Jr, Neet K. E. The catalytic and regulatory properties of enzymes. Annu Rev Biochem. 1968;37:359–410. doi: 10.1146/annurev.bi.37.070168.002043. [DOI] [PubMed] [Google Scholar]
  9. Medzihradzky K., Voynick I. M., Medzihradszky-Schweiger H., Fruton J. S. Effect of secondary enzyme-substrate interactions on the cleavage of synthetic peptides by pepsin. Biochemistry. 1970 Mar 3;9(5):1154–1162. doi: 10.1021/bi00807a016. [DOI] [PubMed] [Google Scholar]
  10. Morávek L., Kostka V. Tentative amino acid sequence of hog pepsin. FEBS Lett. 1973 Sep 15;35(2):276–278. doi: 10.1016/0014-5793(73)80303-5. [DOI] [PubMed] [Google Scholar]
  11. Pedersen V. B., Foltmann B. The amino acid sequence of the first 61 residues of chymosin (rennin EC 3.4.4.3). FEBS Lett. 1973 Sep 15;35(2):250–254. doi: 10.1016/0014-5793(73)80297-2. [DOI] [PubMed] [Google Scholar]
  12. Raju E. V., Humphreys R. E., Fruton J. S. Gel filtration studies on the binding of peptides to pepsin. Biochemistry. 1972 Sep 12;11(19):3533–3536. doi: 10.1021/bi00769a007. [DOI] [PubMed] [Google Scholar]
  13. Sachdev G. P., Brownstein A. D., Fruton J. S. N-methyl-2-anilinonaphthalene-6-sulfonyl peptides as fluorescent probes for pepsin-substrate interaction. J Biol Chem. 1973 Sep 25;248(18):6292–6299. [PubMed] [Google Scholar]
  14. Sachdev G. P., Fruton J. S. Pyridyl esters of peptides as synthetic substrates of pepsin. Biochemistry. 1969 Nov;8(11):4231–4238. doi: 10.1021/bi00839a002. [DOI] [PubMed] [Google Scholar]
  15. Sachdev G. P., Fruton J. S. Secondary enzyme-substrate interactions and the specificity of pepsin. Biochemistry. 1970 Nov 10;9(23):4465–4470. doi: 10.1021/bi00825a001. [DOI] [PubMed] [Google Scholar]
  16. Tang J., Sepulveda P., Marciniszyn J., Jr, Chen K. C., Huang W. Y., Tao N., Liu D., Lanier J. P. Amino-acid sequence of porcine pepsin. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3437–3439. doi: 10.1073/pnas.70.12.3437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Thompson R. C., Blout E. R. Dependence of the kinetic parameters for elastase-catalyzed amide hydrolysis on the length of peptide substrates. Biochemistry. 1973 Jan 2;12(1):57–65. doi: 10.1021/bi00725a011. [DOI] [PubMed] [Google Scholar]
  18. Voynick I. M., Fruton J. S. The comparative specificity of acid proteinases. Proc Natl Acad Sci U S A. 1971 Feb;68(2):257–259. doi: 10.1073/pnas.68.2.257. [DOI] [PMC free article] [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