A Modified Arrhenius Approach to Thermodynamically Study Regioselectivity in Cytochrome P450-Catalyzed Substrate Conversion

Rosa A. Luirink, Marlies C.A. Verkade-Vreeker, Jan N.M. Commandeur, Daan P. Geerke*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

The regio- (and stereo-)selectivity and specific activity of cytochrome P450s are determined by the accessibility of potential sites of metabolism (SOMs) of the bound substrate relative to the heme, and the activation barrier of the regioselective oxidation reaction(s). The accessibility of potential SOMs depends on the relative binding free energy (ΔΔGbind) of the catalytically active substrate-binding poses, and the probability of the substrate to adopt a transition-state geometry. An established experimental method to measure activation energies of enzymatic reactions is the analysis of reaction rate constants at different temperatures and the construction of Arrhenius plots. This is a challenge for multistep P450-catalyzed processes that involve redox partners. We introduce a modified Arrhenius approach to overcome the limitations in studying P450 selectivity, which can be applied in multiproduct enzyme catalysis. Our approach gives combined information on relative activation energies, ΔΔGbind values, and collision entropies, yielding direct insight into the basis of selectivity in substrate conversion.

Original languageEnglish
Pages (from-to)1461-1472
Number of pages12
JournalChemBioChem
Volume21
Issue number10
Early online date9 Jan 2020
DOIs
Publication statusPublished - 15 May 2020

Keywords

  • cytochrome P450
  • molecular dynamics
  • selectivity
  • thermodynamics
  • validation

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