title: A study of glycerolphosphate acyltransferase in rat liver mitochondria-submitochondrial localization and some properties of the solubilized enzyme.
abstract: 1. Glycerolphosphate acyltransferase (GPAT) was solubilized from the rat liver mitochondrial membranes using sodium cholate. Dithiothreitol was necessary to stabilize the solubilized enzyme on storage. 2. Unlike the enzyme in situ in mitochondrial membranes, the solubilized mitochondrial GPAT was susceptible to inhibition by N-ethylmaleimide; a property more characteristic of the distinct microsomal form of GPAT. 3. Solubilized mitochondrial GPAT retained its very high preference for saturated acyl-CoA substrate (palmitoyl-CoA) and had no activity whatever with any tested concentration of the unsaturated substrate oleoyl-CoA. 4. Solubilization increased the affinity of mitochondrial GPAT for palmitoyl-CoA whilst decreasing the Km for glycerol phosphate. 5. After separation of liver mitochondrial outer and inner membranes and estimation of cross-contamination by appropriate markers it was concluded that the mitochondrial inner membrane contains significant GPAT activity. This was established with preparations from fed, 48 hr-starved and streptozotocin-diabetic rats.
authors.affiliation: Department of Biochemistry & Molecular Biology, University College London, U.K.
authors.forename: J R
authors.initials: JR
authors.lastname: Mitchell
authors.forename: E D
authors.initials: ED
authors.lastname: Saggerson
mesh_terms: D000818:Animals; D004795:Enzyme Stability; D005033:Ethylmaleimide; D005992:Glycerol-3-Phosphate O-Acyltransferase / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000737:chemistry; D007425:Intracellular Membranes / Q000201:enzymology*; D008297:Male; D008930:Mitochondria, Liver / Q000201:enzymology* / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D012995:Solubility
chemical_list: D005992:Glycerol-3-Phosphate O-Acyltransferase; D005033:Ethylmaleimide