FORMATION OF A PORPHYRIN PI-CATION RADICAL IN THE FLUORIDE COMPLEX OF HORSERADISH-PEROXIDASE

Horseradish peroxidase (HRP) was oxidized by IrCl62- to a mixture of compounds I and II, the rate of oxidation and the ratio of-the mixture being greatly affected by pH (Hayashi & Yamazaki, 1979). Oxidation of HRP by IrCl62- in the presence of fluoride was significantly accelerated. This resulted in the formation of a new compound which is a ferric fluoride complex containing a porphyrin pi-cation radical. The spectrum of the new compound showed a decreased absorption band in the Sorer region and a broad band at 570 nm; which was converted to that of the original ferric fluoride complex by addition of ascorbate or hydroquinone. Addition of cyanide slowed down the oxidation of HRP by IrCl62-, and the oxidation product was the same as that obtained in the absence of cyanide. Compound I was formed when H2O2 was added to HRP in the presence of fluoride or cyanide. The one-electron reduction potential (E(o)') of the oxidized HRP-fluoride complex was measured at several pH values, the E(o)' value at pH 7 being 861 +/- 4 mV. The ratio of Delta E(o)' to Delta pH was 49 mV/pH unit.