Magnetic spin effects in enzymatic reactions: radical oxidation of NADH by horseradish peroxidase

J Am Chem Soc. 2006 Jul 5;128(26):8651-8. doi: 10.1021/ja0585735.

Abstract

A description of the elementary steps of the horseradish peroxidase (HRP)-catalyzed oxidation of NADH is presented, along with a quantitative analysis of the magnetic-field dependence of the enzymatic reaction. In the absence of H(2)O(2), the catalytic cycle begins with single-electron transfer from NADH to native HRP to form the NADH(.+) radical cation and the ferroperoxidase intermediate (Per(2+)). The theoretical framework for the magnetic-field dependent recombination of radical pairs has been extended to describe the magnetic-field dependence of reaction rate constants for multi-spin paramagnetic pairs, including the NADH(.+) radical cation and Per(2+) that exist in a correlated quartet electronic spin state. Good agreement between the experimentally observed and the theoretically calculated magnetic-field dependences of the effective rate constants underlines the importance of the initial single-electron-transfer step and supports a model in which the catalytic cycle begins with the one-electron reduction of HRP by NADH.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalysis
  • Enzyme Activation
  • Free Radicals / chemical synthesis
  • Free Radicals / chemistry
  • Horseradish Peroxidase / chemistry*
  • Kinetics
  • Magnetics*
  • Models, Chemical
  • NAD / chemistry*
  • Oxidation-Reduction
  • Time Factors

Substances

  • Free Radicals
  • NAD
  • Horseradish Peroxidase