High-valent intermediates in the reaction of N alpha-acetyl microperoxidase-8 with hydrogen peroxide: models for compounds 0, I and 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 degrees 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 kmaxobs = 95 s-1 and KM = 87 mM. Rapid-scan studies carried out with [H2O2] = 200 mM at -38.0 degrees 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.
Wang
J S
JS
Department of Chemistry, Florida State University, Tallahassee 32306.
Baek
H K
HK
Van Wart
H E
HE
D000818:Animals; D003574:Cytochrome c Group / Q000378:metabolism*; D006420:Hemeproteins / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism*; D006736:Horses; D006861:Hydrogen Peroxide / Q000378:metabolism*; D007700:Kinetics; D008433:Mathematics; D008962:Models, Theoretical; D010446:Peptide Fragments / Q000378:metabolism*; D013053:Spectrophotometry; D013059:Spectrum Analysis, Raman
D003574:Cytochrome c Group; D006420:Hemeproteins; D010446:Peptide Fragments; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; C071067:N(alpha)-acetylmicroperoxidase-8