Nitric oxide-mediated inactivation of mammalian ferrochelatase in vivo and in vitro: possible involvement of the iron-sulphur cluster of the enzyme.
To investigate the role of the iron-sulphur cluster in mammalian ferrochelatases, the terminal enzyme of the haem biosynthetic pathway, we examined the interaction of nitric oxide (NO) and ferrochelatase. When macrophage cell line RAW 264.7 cells were treated with interferon-gamma and lipopolysaccharide NO synthesis in the cells was stimulated, and a decrease in ferrochelatase activity was observed, with no change in the amount of ferrochelatase. The addition of NG-monomethyl-L-arginine, a selective inhibitor of NO synthesis, reduced the effect of interferon-gamma and lipopolysaccharide, while the effect of NG-monomethyl-L-arginine was suppressed by the addition of L-arginine, a substrate of NO synthase. When purified recombinant human ferrochelatase was treated with 3-morpholinosydnonimine, a NO-generating compound, ferrochelatase activity decreased with disappearance of characteristic absorbance spectra of the iron-sulphur cluster. S-Nitroso-N-acetylpenicillamine also reduced the activity, in a dose-dependent manner. These results indicate that ferrochelatase activity can be modulated by NO synthesis probably through disruption of the iron-sulphur cluster. We propose that inactivation of ferrochelatase mediated by NO (or NO-derived species) may play a role in the regulation of haem metabolism.
Furukawa
T
T
Department of Hygiene, Kansai Medical University, Osaka, Japan.
Kohno
H
H
Tokunaga
R
R
Taketani
S
S
D000594:Amino Acid Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D002460:Cell Line; D003001:Cloning, Molecular; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli; D005294:Ferrochelatase / Q000037:antagonists & inhibitors* / Q000737:chemistry; D006801:Humans; D015151:Immunoblotting; D007371:Interferon-gamma / Q000494:pharmacology*; D007506:Iron-Sulfur Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages; D008322:Mammals; D051379:Mice; D008981:Molsidomine / Q000031:analogs & derivatives / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis / Q000502:physiology*; D019001:Nitric Oxide Synthase; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors; D026423:S-Nitroso-N-Acetylpenicillamine; D014665:Vasodilator Agents / Q000494:pharmacology; D019323:omega-N-Methylarginine
D004791:Enzyme Inhibitors; D007506:Iron-Sulfur Proteins; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D014665:Vasodilator Agents; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; C002385:linsidomine; D026423:S-Nitroso-N-Acetylpenicillamine; D007371:Interferon-gamma; D001120:Arginine; D008981:Molsidomine; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; D005294:Ferrochelatase; D010396:Penicillamine