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
. 1969 Aug;63(4):1214–1221. doi: 10.1073/pnas.63.4.1214

ALKALINE PH DEPENDENCE OF δCHYMOTRYPSIN-CATALYZED HYDROLYSIS OF SPECIFIC SUBSTRATES*

Pablo Valenzuela 1,†, Myron L Bender 1
PMCID: PMC223452  PMID: 5260922

Abstract

Km (app) and kcat for the δchymotrypsin-catalyzed hydrolysis of N-acetyl-L-tryptophan methyl ester and N-furylacryloyl-L-tryptophanamide were measured as a function of pH and ionic strength. The Km (app) values do not increase considerably above pH 9 for δ-chymotrypsin, as is the case with α-chymotrypsin. The observed kinetic difference between both enzymes at high pH suggests that the reversible inactivation of α-chymotrypsin at alkaline pH may involve the participation of tyrosine 146 or alanine 149 since both residues are present as chain termini in α-chymotrypsin but not in δ-chymotrypsin.

Full text

PDF
1214

Selected References

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

  1. Bender M. L., Gibian M. J., Whelan D. J. The alkaline pH dependence of chymotrypsin reactions: postulation of a pH-dependent intramolecular competitive inhibition. Proc Natl Acad Sci U S A. 1966 Sep;56(3):833–839. doi: 10.1073/pnas.56.3.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brandt K. G., Himoe A., Hess G. P. Investigations of the chymotrypsin-catalyzed hydrolysis of specific substrates. 3. Determination of individual rate constants and enzyme-substrate binding constants for specific amide and ester substrates. J Biol Chem. 1967 Sep 10;242(17):3973–3982. [PubMed] [Google Scholar]
  3. DIXON M. The effect of pH on the affinities of enzymes for substrates and inhibitors. Biochem J. 1953 Aug;55(1):161–170. doi: 10.1042/bj0550161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. GLADNER J. A., NEURATH H. Carboxyl terminal groups of proteolytic enzymes. II. Chymotrypsins. J Biol Chem. 1954 Feb;206(2):911–924. [PubMed] [Google Scholar]
  5. Glazer A. N., Sanger F. The iodination of chymotrypsinogen. Biochem J. 1964 Jan;90(1):92–98. doi: 10.1042/bj0900092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Himoe A., Hess G. P. On the elucidation of the pH dependence of chymotrypsin catalyzed reactions at alkaline pH. Biochem Biophys Res Commun. 1966 May 3;23(3):234–239. doi: 10.1016/0006-291x(66)90533-x. [DOI] [PubMed] [Google Scholar]
  7. Himoe A., Parks P. C., Hess G. P. Investigations of the chymotrypsin-catalyzed hydrolysis of specific substrates. I. The pH dependence of the catalytic hydrolysis of N-acetyl-L-tryptophanamide by three forms of the enzyme at alkaline pH. J Biol Chem. 1967 Mar 10;242(5):919–929. [PubMed] [Google Scholar]
  8. Labouesse J., Gervais M. Preparation of chemically defined epsilon N-acetylated trypsin. Eur J Biochem. 1967 Sep;2(2):215–223. doi: 10.1111/j.1432-1033.1967.tb00127.x. [DOI] [PubMed] [Google Scholar]
  9. McConn J., Fasman G. D., Hess G. P. Conformation of the high pH form of chymotrypsin. J Mol Biol. 1969 Feb 14;39(3):551–562. doi: 10.1016/0022-2836(69)90145-4. [DOI] [PubMed] [Google Scholar]
  10. McConn J., Ku E., Odell C., Czerlinski G., Hess G. P. Conformation and activity of chymotrypsin: the pH-dependent, substrate-induced proton uptake. Science. 1968 Jul 19;161(3838):274–276. doi: 10.1126/science.161.3838.274. [DOI] [PubMed] [Google Scholar]
  11. Oppenheimer H. L., Labouesse B., Hess G. P. Implication of an ionizing group in the control of conformation and activity of chymotrypsin. J Biol Chem. 1966 Jun 10;241(11):2720–2730. [PubMed] [Google Scholar]
  12. SCHONBAUM G. R., ZERNER B., BENDER M. L. The spectrophotometric determination of the operational normality of an alpha-chymotrypsin solution. J Biol Chem. 1961 Nov;236:2930–2935. [PubMed] [Google Scholar]
  13. Sanger F. The free amino groups of insulin. Biochem J. 1945;39(5):507–515. doi: 10.1042/bj0390507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sigler P. B., Blow D. M., Matthews B. W., Henderson R. Structure of crystalline -chymotrypsin. II. A preliminary report including a hypothesis for the activation mechanism. J Mol Biol. 1968 Jul 14;35(1):143–164. doi: 10.1016/s0022-2836(68)80043-9. [DOI] [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