Characterization of the enzymatic and nonenzymatic peroxidative degradation of iron porphyrins and cytochrome P-450 heme

Biochemistry. 1985 Jun 18;24(13):3254-63. doi: 10.1021/bi00334a027.

Abstract

Both purified cytochrome P-450 (P-450) and free ferriprotoporphyrin IX are destroyed by NADPH-P-450 reductase in the presence of NADPH and O2. The process appears to be mediated by H2O2 generated by reduction of O2. Six major products were identified from the reaction of H2O2 with ferri-protoporphyrin IX-hematinic acid, methylvinylmaleimide, and four dipyrrolic propentdyopents. The structures of the propentdyopents were elucidated by mass spectrometry and 1H NMR methods. Both free ferriprotoporphyrin IX and P-450 yielded these same products in similar relative ratios. P-450 heme in rat liver microsomes was degraded in the presence of O2 and NADPH and either NaN3 (a catalase inhibitor) or Fe-ADP (which promotes lipid peroxidation); the products were primarily hematinic acid, methylvinylmaleimide, and small quantities of one propentdyopent. Only the two maleimides were detected in the destruction of microsomal P-450 heme by cumene hydroperoxide and iodosylbenzene. On the basis of the reaction of H2O2 with several metal-octaethylethylporphyrin complexes and free octaethylporphyrin, the iron chelated in ferriprotoporphyrin IX is required for degradation by H2O2. Biliverdin is not an intermediate in the formation of maleimides and propentdyopents from heme. Experiments using the tetraethylpropentdyopent produced from ferrioctaethylporphyrin suggest that propentdyopents are not further cleaved to form the maleimides. A mechanism for oxidative heme destruction consistent with these observations is proposed.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Bilirubin / metabolism
  • Biliverdine / metabolism
  • Cytochrome P-450 Enzyme System / metabolism*
  • Heme / metabolism*
  • Hydrogen Peroxide
  • Magnetic Resonance Spectroscopy
  • Male
  • Microsomes, Liver / enzymology
  • NADPH-Ferrihemoprotein Reductase / metabolism*
  • Porphyrins / chemical synthesis
  • Porphyrins / metabolism*
  • Rats
  • Rats, Inbred Strains

Substances

  • Porphyrins
  • Heme
  • Cytochrome P-450 Enzyme System
  • Hydrogen Peroxide
  • NADPH-Ferrihemoprotein Reductase
  • Biliverdine
  • Bilirubin