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A traffic light enzyme: acetate binding reversibly switches chlorite dismutase from a red- to a green-colored heme protein.
JBIC Journal of Biological Inorganic Chemistry ( IF 2.7 ) Pub Date : 2020-04-03 , DOI: 10.1007/s00775-020-01784-1 Durga Mahor 1 , Julia Püschmann 1 , Menno van den Haak 1 , Pepijn J Kooij 1 , David L J van den Ouden 1 , Marc J F Strampraad 1 , Batoul Srour 1, 2 , Peter-Leon Hagedoorn 1
中文翻译:
一个交通信号灯酶:乙酸酯结合可逆地将亚氯酸盐歧化酶从红色变为绿色的血红素蛋白。
更新日期:2020-04-03
JBIC Journal of Biological Inorganic Chemistry ( IF 2.7 ) Pub Date : 2020-04-03 , DOI: 10.1007/s00775-020-01784-1 Durga Mahor 1 , Julia Püschmann 1 , Menno van den Haak 1 , Pepijn J Kooij 1 , David L J van den Ouden 1 , Marc J F Strampraad 1 , Batoul Srour 1, 2 , Peter-Leon Hagedoorn 1
Affiliation
Abstract
Chlorite 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
中文翻译:
一个交通信号灯酶:乙酸酯结合可逆地将亚氯酸盐歧化酶从红色变为绿色的血红素蛋白。