Citation

Hiner AN, Hernández-Ruiz J, Rodríguez-López JN, Arnao MB, Varon R, García-Cánovas F, Acosta M. Journal of biological inorganic chemistry: JBIC: a publication of the Society of Biological Inorgani 2001; 6(5-6): 504-516.

Copyright

(Copyright © 2001)

DOI

10.1007/s007750100219

PMID

11472014

Abstract

The inactivation of horseradish peroxidase A2 (HRP-A2) with H2O2 as the sole substrate has been studied. In incubation experiments it was found that the fall in HRP-A2 activity was non-linearly dependent on H2O2 concentrations and that a maximum level of inactivation of approximately 80% (i.e. approximately 20% residual activity) was obtained with 2,000 or more equivalents of H2O2. Further inactivation was only induced at much higher H2O2 concentrations. Spectral changes during incubations of up to 5 days showed the presence of a compound III-like species whose abundance was correlated to the level of resistance observed. Inactivation was pH dependent, the enzyme being much more sensitive under acid conditions. A partition ratio (r1 approximately equals 1,140 at pH 6.5) between inactivation and catalysis was calculated from the data. The kinetics of inactivation followed single exponential time curves and were H2O2 concentration dependent. The apparent maximum rate constant of inactivation was lambdamax=3.56+/-0.07x10(-4)s(-1) and the H2O2 concentration required to give lambdamax/2 was K2=9.94+/-0.52 mM. The relationship lambdamax

Language: en

Keywords

Catalase; Enzyme Activation; Enzyme Stability; Horseradish Peroxidase; Hydrogen Peroxide; Isoenzymes; Kinetics; Oxidants; Oxidative Stress; Tetranitromethane; Time Factors