Differential scanning calorimetric study of carboxypeptidase B, procarboxypeptidase B and its globular activation domain

Eur J Biochem. 1991 Sep 15;200(3):663-70. doi: 10.1111/j.1432-1033.1991.tb16230.x.

Abstract

High-sensitivity differential scanning calorimetry has been applied to the study of porcine pancreatic carboxypeptidase B, the proenzyme and its 81-residue activation domain. The thermal study has been carried out over a range of scan rates, ionic strengths and pH values. The thermal unfolding of the isolated activation domain has been found to be reversible and corresponds to that of a typical compact globular structure, with melting temperatures higher than those of the enzyme and proenzyme. Both proteins, on the other hand, undergo an irreversible, highly scan-rate-dependent thermal denaturation under all the experimental conditions investigated. The denaturation of the enzyme at pH 7.5 and the proenzyme at pH 7.5 and 9.0 follows the two-state irreversible model [Sánchez-Ruiz, J.M., López-Lacomba, J.L., Cortijo, M. & Mateo, P.L. (1988) Biochemistry 27, 1648-1652]. Thus the kinetic constants and activation parameters of the denaturation process could be obtained and compared to those for other proteins, particularly those of the closely related carboxypeptidase A system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calorimetry, Differential Scanning*
  • Carboxypeptidase B
  • Carboxypeptidases / chemistry*
  • Enzyme Activation
  • Enzyme Precursors / chemistry*
  • Hot Temperature
  • Hydrogen-Ion Concentration
  • Osmolar Concentration
  • Pancreas / enzymology*
  • Protein Conformation
  • Protein Denaturation
  • Swine
  • Thermodynamics

Substances

  • Enzyme Precursors
  • Carboxypeptidases
  • Carboxypeptidase B