A radiochemical assay for heme synthase activity.
A radiochemical assay for heme synthase (ferrochelatase) activity is described in this report. The principle of the assay is to measure the incorporation of 59Fe, added to the reaction flask with a known amount of iron, into heme. Iron is maintained in the ferrous state, and oxygen is excluded from the flask during the incubation of the substrates with enzyme. Labeled heme product is extracted by solvent partitioning and counted in a gamma scintillation spectrometer. The assay can detect picomole amounts of product and is most useful when low levels of heme synthase activity are present.
D000818:Animals; D005294:Ferrochelatase / Q000032:analysis*; D005347:Fibroblasts; D005615:Freezing; D066298:In Vitro Techniques; D007501:Iron / Q000378:metabolism; D007504:Iron Radioisotopes; D008099:Liver / Q000378:metabolism; D008190:Lyases / Q000032:analysis*; D008297:Male; D008930:Mitochondria, Liver / Q000201:enzymology; D011166:Porphyrins / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D007504:Iron Radioisotopes; D011166:Porphyrins; D007501:Iron; D008190:Lyases; D005294:Ferrochelatase