tailieunhanh - Báo cáo Y học: Spectroscopic characterization and ligand-binding properties of chlorite dismutase from the chlorate respiring bacterial strain GR-1

Chlorite dismutase (EC ), an enzyme capable of reducing chlorite to chloride while producing molecular oxygen, has been characterized using EPR and optical spectroscopy. The EPR spectrum of GR-1 chlorite dis-mutase shows two different high-spin ferric heme species, whichwe have designated narrow (gx,y,z¼, , ) and broad (gz,y,x ¼, , ). | Eur. J. Biochem. 269 4905-4911 2002 FEBS 2002 doi Spectroscopic characterization and ligand-binding properties of chlorite dismutase from the chlorate respiring bacterial strain GR-1 Peter L. Hagedoorn Daniel C. de Geus and Wilfred R. Hagen Kluyver Department of Biotechnology Delft University of Technology The Netherlands Chlorite dismutase EC an enzyme capable of reducing chlorite to chloride while producing molecular oxygen has been characterized using EPR and optical spectroscopy. The EPR spectrum of GR-1 chlorite dismutase shows two different high-spin ferric heme species which we have designated narrow gx y z and broad gz y x . Spectroscopic evidence is presented for a proximal histidine co-ordinating the heme iron center of the enzyme. The UV visible spectrum of the ferrous enzyme and EPR spectra of the ferric hydroxide and imidazole adducts are characteristic of a heme protein with an axial histidine co-ordinating the iron. Furthermore the substrate analogs nitrite and hydrogen peroxide have been found to bind to ferric chlorite dismutase. EPR spectroscopy of the hydrogen peroxide adduct shows the loss of both high-spin and low-spin ferric signals and the appearance of a sharp radical signal. The NO adduct of the ferrous enzyme exhibits a low-spin EPR signal typical of a five-co-ordinate heme iron nitrosyl adduct. It seems that the bond between the proximal histidine and the iron is weak and can be broken upon binding of NO. The midpoint potential Em Fe3 2 -23 mV of chlorite dismutase is higher than for most heme enzymes. The spectroscopic features and redoxproperties of chlorite dismutase are more similar to the gas-sensing hemoproteins such as guanylate cyclase and the globins than to the heme enzymes. Keywords chlorate respiration chlorite dismutase EPR ESR heme enzyme. Chlorate and chlorite are degradation products of the commonly used bleaching agent chlorine dioxide. Recently .

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