A rapid assay of acyl-coenzyme A:lysolecithin acyltransferase activity.
A simple and rapid procedure for the assay of acyl-coenzyme A:1-acyl-sn-glycero-3-phosphocholine acyltransferase (lysolecithin acyltransferase, LLAT [EC 2.3.1.23]) activity in crude enzyme preparations is described. The incubation system utilizes lysolecithin and [1-14C]-oleoyl-coenzyme A as substrates. Labeled fatty acid released due to accompanying acyl-coenzyme A hydrolase [EC 3.1.2.2]activity is first removed by di-isopropyl ether extraction. The labeled lecithin produced due to LLAT action is then quantitatively recovered by partition of the incubation medium with di-isopropyl ether-n-butanol 60:40 (v/v). Selective extraction of the labeled lecithin formed and avoidance of customary thin-layer chromatographic isolation procedures permits assay of LLAT activity with excellent accuracy at a substantial saving of time. The entire assay can be completed in less than 30 min as compared to 2-3 hrs when following conventional procedures.
D008243:1-Acylglycerophosphocholine O-Acyltransferase / Q000032:analysis*; D000217:Acyltransferases / Q000032:analysis*; D000818:Animals; D003065:Coenzyme A; D005227:Fatty Acids / Q000032:analysis; D008244:Lysophosphatidylcholines; D008722:Methods; D008862:Microsomes, Liver / Q000201:enzymology; D010170:Palmitoyl-CoA Hydrolase / Q000032:analysis; D010713:Phosphatidylcholines / Q000032:analysis; D051381:Rats
D005227:Fatty Acids; D008244:Lysophosphatidylcholines; D010713:Phosphatidylcholines; D000217:Acyltransferases; D008243:1-Acylglycerophosphocholine O-Acyltransferase; D010170:Palmitoyl-CoA Hydrolase; D003065:Coenzyme A