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A traffic light enzyme: acetate binding reversibly switches chlorite dismutase from a red- to a green-colored heme protein

Authors
  • Mahor, Durga1
  • Püschmann, Julia1
  • van den Haak, Menno1
  • Kooij, Pepijn J.1
  • van den Ouden, David L. J.1
  • Strampraad, Marc J. F.1
  • Srour, Batoul1, 2
  • Hagedoorn, Peter-Leon1
  • 1 Delft University of Technology, Van der Maasweg 9, Delft, 2629HZ, The Netherlands , Delft (Netherlands)
  • 2 CEA, CNRS, Univ. Paris‐Sud, Université Paris‐Saclay, Gif‐sur‐Yvette Cedex, 91198, France , Gif‐sur‐Yvette Cedex (France)
Type
Published Article
Journal
JBIC Journal of Biological Inorganic Chemistry
Publisher
Springer-Verlag
Publication Date
Apr 03, 2020
Volume
25
Issue
4
Pages
609–620
Identifiers
DOI: 10.1007/s00775-020-01784-1
Source
Springer Nature
Keywords
License
Green

Abstract

AbstractChlorite dismutase is a unique heme enzyme that catalyzes the conversion of chlorite to chloride and molecular oxygen. The enzyme is highly specific for chlorite but has been known to bind several anionic and neutral ligands to the heme iron. In a pH study, the enzyme changed color from red to green in acetate buffer pH 5.0. The cause of this color change was uncovered using UV–visible and EPR spectroscopy. Chlorite dismutase in the presence of acetate showed a change of the UV–visible spectrum: a redshift and hyperchromicity of the Soret band from 391 to 404 nm and a blueshift of the charge transfer band CT1 from 647 to 626 nm. Equilibrium binding titrations with acetate resulted in a dissociation constant of circa 20 mM at pH 5.0 and 5.8. EPR spectroscopy showed that the acetate bound form of the enzyme remained high spin S = 5/2, however with an apparent change of the rhombicity and line broadening of the spectrum. Mutagenesis of the proximal arginine Arg183 to alanine resulted in the loss of the ability to bind acetate. Acetate was discovered as a novel ligand to chlorite dismutase, with evidence of direct binding to the heme iron. The green color is caused by a blueshift of the CT1 band that is characteristic of the high spin ferric state of the enzyme. Any weak field ligand that binds directly to the heme center may show the red to green color change, as was indeed the case for fluoride.Graphic abstract

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