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Kinetic Studies of Immobilized α-Chymotrypsin in Apolar Solvents

  • Myron L. Bender
  • A. B. Cottingham
  • Lee K. Sun
  • K. Tanizawa
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 86A)

Abstract

The mechanism of α-chymotrypsin action has been probed by extending studies of native chymotrypsin to immobilized chymo-trypsin, where the organic content of the solution can be raised to much higher levels and thus one can explicitly look at the role of water. When one does this, one finds that water only appears in the deacylation reaction. The premise that one can go from native chymotrypsin (soluble) to immobilized chymotrypsin (insoluble) has been tested by several criteria. It has been found in many instances that the two are identical: in absolute rate, in pK a. They are, however, not identical to one another in binding, due to differences in diffusion, which is to be expected. Thus, mechanistically immobilized and native chymotrypsin are identical to one another and the use of immobilized chymotrypsin can be used to specify the mechanism even more: it must proceed through two tetrahedral intermediates and two acyl-enzyme intermediates.

Keywords

Ethyl Ester Porous Glass Tetrahedral Intermediate Apolar Solvent Zole Group 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Bender, M. L., Clement, G. E., Gunter, C. R., and Kezdy, F. J. (1964). The Kinetics of α-Chymotrypsin Reactions in the presence of added Nucleophiles. J. Amer. Chem, Soc., 86, 3697.CrossRefGoogle Scholar
  2. Bender, M. L. and Kezdy, F. J. (1965). Mechanism of Action of Proteolytic Enzymes. Ann. Rev. Biochem., 34, 49.PubMedCrossRefGoogle Scholar
  3. Bundy, H. F. (1962). A New Spectrophotometric Method for the Determination of Chymotrypsin Activity. Anal. Biochem., 3, 431.PubMedCrossRefGoogle Scholar
  4. Kezdy, F. J., Clement, G. E., and Bender, M. L. (1964). The Observation of Acyl-Enzyme Intermediates in the Chymotrypsin-catalyzed Reactions of N-acetyl-L-tryptophan Derivatives at low pH. J. Amer. Chem. Soc., 86, 3690.CrossRefGoogle Scholar
  5. Komiyama, M., Breaux, E.J., and Bender, M. L. (1976). Cyclo-amylose-catalyzed Hydrolysis as a Model for the “Charge-Relay” System. Submitted to Bioorganic Chemistry.Google Scholar
  6. Schonbaum, G. R., Zerner, B, and Bender, M. L. (1960). The Spectrophotometric Determination of the Operational Normality of an α-Chymotrypsin Solution. J. Biol. Chem., 235, 2930.Google Scholar

Copyright information

© Plenum Press, New York 1977

Authors and Affiliations

  • Myron L. Bender
    • 1
  • A. B. Cottingham
    • 1
  • Lee K. Sun
    • 1
  • K. Tanizawa
    • 1
  1. 1.Division of Biochemistry, Dept. of ChemistryNorthwestern UniversityEvanstonUSA

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