HIGH-VALENT INTERMEDIATES IN THE REACTION OF N-ALPHA-ACETYL MICROPEROXIDASE-8 WITH HYDROGEN-PEROXIDE - MODELS FOR COMPOUNDS-0, COMPOUNDS-I AND COMPOUNDS-II OF HORSERADISH-PEROXIDASE

N-acetyl microperoxidase-8 (Ac-MP-8) is a water soluble, ferric heme model for the peroxidases. The reaction of Ac-MP-8 with H2O2 in 10 mM potassium phosphate over the pH range of 7-11 gives rise sequentially to relatively stable green and red species with properties that closely mimic those of HRP compounds I and II, respectively. Low-temperature stopped-flow studies of this reaction carried out in 50% v/v methanol/10 mM potassium phosphate, pH* 9.1 at -25.8°C indicate that the pseudo-first-order rate constant, kobs, that describes the formation of the green intermediate exhibits saturation kinetics as a function of [H2O2] with kobsmax = 95 s-1 and KM = 87 mM. Rapid-scan studies carried out with [H2O2] = 200 mM at -38.0°C show that a compound 0 species with a characteristic band near 340 nm is formed whose conversion to the green species is rate limiting. Thus, Ac-MP-8 has high-valent forms that are models for all three known intermediates in the peroxidase cycle of horseradish peroxidase. © 1991.