Effect of lipids and organic solvents on the enzymic formation of zinc protoporphyrin and haem.
1. Differences observed in earlier work between the enzymic chelation with protoporphyrin of Zn(2+) and Fe(2+) ions respectively have now been explained as being caused by the presence of peroxides in the ether used in the enzyme assay. The inhibitory effect of peroxides is established by the reducing agent, which is present in the assay for chelation of iron but not in that for zinc. There are now no reasons for the belief that two different enzymes catalyse formation of complexes with zinc and iron respectively. 2. Removal of lipid from both chromatophores and mitochondria markedly reduced chelatase activity. Activity could be partially restored by the addition of lipid fractions. Phosphatidic acid, but not phosphatidylcholine or phosphatidylethanolamine, actively stimulated the formation of zinc protoporphyrin and haem by chromatophores and mitochondrial preparations. 3. Lipid-containing extracts of chromatophores, and fractions thereof obtained by silicic acid chromatography, partially restored chelatase activity of Tween extracts of mitochondria. Thus, although both enzymes are considered to be lipoproteins, the identity of the lipids concerned is still uncertain. 4. A great number of organic solvents such as esters, ethers, ketones and, to a lesser extent, alcohols, stimulate enzymic chelation of both metals with protoporphyrin. A number of explanations for these findings are considered and it is suggested that organic solvents interact in some way with the enzyme lipoprotein, changing either its conformation or allowing closer contact between the enzyme and its substrates.
D000085:Acetates / Q000494:pharmacology; D000438:Alcohols / Q000494:pharmacology; D020130:Bacterial Chromatophores / Q000201:enzymology / Q000378:metabolism*; D004147:Dioxins / Q000494:pharmacology; D004987:Ethers / Q000494:pharmacology; D005561:Formates / Q000494:pharmacology; D006418:Heme / Q000096:biosynthesis*; D066298:In Vitro Techniques; D007659:Ketones / Q000494:pharmacology; D008314:Malonates / Q000494:pharmacology; D008928:Mitochondria / Q000201:enzymology; D010743:Phospholipids / Q000494:pharmacology*; D011166:Porphyrins / Q000096:biosynthesis*; D012241:Rhodopseudomonas / Q000378:metabolism*; D012997:Solvents / Q000494:pharmacology*
D000085:Acetates; D000438:Alcohols; D004147:Dioxins; D004987:Ethers; D005561:Formates; D007659:Ketones; D008314:Malonates; D010743:Phospholipids; D011166:Porphyrins; D012997:Solvents; D006418:Heme