Partial purification of diphosphatidylglycerol synthetase from liver mitochondrial membranes.
The enzyme responsible for the conversion of phosphatidylglycerol to diphosphatidylglycerol (cardiolipin) in the presence of cytidine diphosphate diacylglycerol is firmly associated with mitochondrial membranes and is not extracted with hypotonic or hypertonic media or with nonionic detergents. Some solubilization was obtained with bile salt solutions, but the zwitter-ionic detergent. Miranol H2M, was most effective in extracting the enzyme. The Miranol extracts were fractionated by column chromatography on Bio-Gel A-1.5 m. The solubilized enzyme is considerably more active in converting unsaturated than saturated phosphatidyl-glycerols, but shows little preference for the cytidine diphosphate diacylglycerols with different fatty acyl substituents. There is an absolute dependence upon divalent cations with the order of effectiveness: Co2+ much greater than Mn2+ greater than Mg2+. In the presence of optimal levels of Co2+ other divalent cations are inhibitory with the order of inhibition: Cd2+ greater than Zn2+ greater than Ca2+ greater than Ba2+ greater than Cu2+ greater than Hg2+ greater than Ni2+. The solubilized enzyme exhibited no requirement for added phospholipids and several phospholipids inhibited the reaction in the order: diphosphatidylglycerol greater than phosphatidylethanolamine greater than phosphatidylserine greater than phosphatidylinositol.
D000818:Animals; D002413:Cations, Divalent; D007425:Intracellular Membranes / Q000201:enzymology*; D007700:Kinetics; D008565:Membrane Proteins; D008930:Mitochondria, Liver / Q000201:enzymology*; D010743:Phospholipids / Q000494:pharmacology; D010770:Phosphotransferases / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D013329:Structure-Activity Relationship; D013552:Swine; D017855:Transferases (Other Substituted Phosphate Groups)
D002413:Cations, Divalent; D008565:Membrane Proteins; D010743:Phospholipids; D010770:Phosphotransferases; D017855:Transferases (Other Substituted Phosphate Groups); C100091:cardiolipin synthetase