Factors influencing the stability of heme and ferrochelatase: role of oxygen.
Ferrochelatase (EC 4.99.1.1) catalyzed heme synthesis is best accomplished in an anaerobic environment. Factors responsible for this phenomenon are not fully understood. Oxygen sensitivity of this reaction may be due to (a) oxidation of essential thiol groups on the enzyme, (b) oxidation of ferrous ions, or (c) the formation of hydrogen peroxide. These possibilities were investigated using rat liver ferrochelatase preparations and a continuous, dual-wavelength assay. Dithiothreitol and ascorbic acid stimulated the ferrochelatase reaction whereas GSH was not as effective. Addition of GSSG had little influence on the enzyme reaction. Total ferrochelatase activity in the assay remained unaffected at the end of the incubation and inclusion of glutathione peroxidase did not alter these results. Thus, ferrochelatase itself was not inactivated by oxidation. In selenium-deficient rats, the mitochondrial ferrochelatase levels were maintained even when glutathione peroxidase activity was significantly depleted. However, glutathione peroxidase very effectively inhibited the thiol-dependent aerobic degradation of heme. These results suggested that autoxidation of heme and of ferrous ions to the unusable ferric form largely contribute toward the oxygen sensitivity of the ferrochelatase reaction in vitro.
Punekar
N S
NS
Chemistry Department, Indian Institute of Technology, Powai, Bombay.
Gokhale
R S
RS
D000332:Aerobiosis; D000818:Animals; D002374:Catalase / Q000494:pharmacology; D004789:Enzyme Activation; D004795:Enzyme Stability; D005294:Ferrochelatase / Q000737:chemistry*; D005609:Free Radicals; D005979:Glutathione Peroxidase / Q000008:administration & dosage / Q000494:pharmacology; D006418:Heme / Q000737:chemistry*; D006868:Hydrolysis; D008297:Male; D010100:Oxygen / Q000502:physiology*; D051381:Rats; D011919:Rats, Inbred Strains
D005609:Free Radicals; D006418:Heme; D002374:Catalase; D005979:Glutathione Peroxidase; D005294:Ferrochelatase; D010100:Oxygen