Relating Diffusion Along the Substrate Tunnel and Oxygen Sensitivity in Hydrogenase

Overview

Journal Nat Chem Biol

Specialties Biochemistry
Biology
Chemistry

Date 2009 Dec 8

PMID 19966788

Citations 49

Authors

Pierre-Pol Liebgott

Fanny Leroux

Benedicte Burlat

Sebastien Dementin

Carole Baffert

Thomas Lautier

Vincent Fourmond

Pierre Ceccaldi

Christine Cavazza

Isabelle Meynial-Salles

Philippe Soucaille

Juan Carlos Fontecilla-Camps

Bruno Guigliarelli

Patrick Bertrand

Marc Rousset

Christophe Leger

Affiliations

Soon will be listed here.

Abstract

In hydrogenases and many other redox enzymes, the buried active site is connected to the solvent by a molecular channel whose structure may determine the enzyme's selectivity with respect to substrate and inhibitors. The role of these channels has been addressed using crystallography and molecular dynamics, but kinetic data are scarce. Using protein film voltammetry, we determined and then compared the rates of inhibition by CO and O2 in ten NiFe hydrogenase mutants and two FeFe hydrogenases. We found that the rate of inhibition by CO is a good proxy of the rate of diffusion of O2 toward the active site. Modifying amino acids whose side chains point inside the tunnel can slow this rate by orders of magnitude. We quantitatively define the relations between diffusion, the Michaelis constant for H2 and rates of inhibition, and we demonstrate that certain enzymes are slowly inactivated by O2 because access to the active site is slow.

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