Molecular Mechanism of Enzymatic Chlorite Detoxification: Insights from Structural and Kinetic Studies

Overview

Journal ACS Catal

Publisher American Chemical Society

Date 2017 Nov 17

PMID 29142780

Citations 18

Authors

Irene Schaffner

Georg Mlynek

Nicola Flego

Dominic Puhringer

Julian Libiseller-Egger

Leighton Coates

Stefan Hofbauer

Marzia Bellei

Paul G Furtmuller

Gianantonio Battistuzzi

Giulietta Smulevich

Kristina Djinovic-Carugo

Christian Obinger

Affiliations

Soon will be listed here.

Abstract

The heme enzyme chlorite dismutase (Cld) catalyzes the degradation of chlorite to chloride and dioxygen. Although structure and steady-state kinetics of Clds have been elucidated, many questions remain (e.g., the mechanism of chlorite cleavage and the pH dependence of the reaction). Here, we present high-resolution X-ray crystal structures of a dimeric Cld at pH 6.5 and 8.5, its fluoride and isothiocyanate complexes and the neutron structure at pH 9.0 together with the pH dependence of the Fe(III)/Fe(II) couple, and the UV-vis and resonance Raman spectral features. We demonstrate that the distal Arg127 cannot act as proton acceptor and is fully ionized even at pH 9.0 ruling out its proposed role in dictating the pH dependence of chlorite degradation. Stopped-flow studies show that (i) Compound I and hypochlorite do not recombine and (ii) Compound II is the immediately formed redox intermediate that dominates during turnover. Homolytic cleavage of chlorite is proposed.

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