Isolation and properties of porcine lecithin:cholesterol acyltransferase.
Lecithin: cholesterol acyltransferase (LCAT, phosphatidylcholine: sterol O-acyltransferase, EC 2.3.1.43) was purified approximately 20 000-fold from pig plasma by ultracentrifugation, phenyl-Sepharose and hydroxyapatite chromatography. Purified LCAT had an apparent relative molecular mass of 69 000 +/- 2000. By isoelectrofocusing it separated into five or six bands with pI values ranging from pH 4.9 to 5.2. The amino acid composition was similar to that of the human enzyme. An antibody against pig LCAT was prepared in goat. The antibody reacted against pig LCAT and gave a reaction of partial identity with human LCAT. Incubation of pig plasma or purified enzyme with the antibody virtually inhibited LCAT activity. The same amount of antibody inactivated only 62% of the LCAT activity in human serum. Pig and human LCAT were activated to the same extent by either human or pig apolipoprotein A-I (apo-A-I) using small liposomes as substrate. Human apoA-I, however, caused a higher esterification rate for both enzymes. Using apoA-I and small liposomes as a substrate, the addition of apoC-II up to 4 micrograms/ml had no effect on the LCAT reaction, but above this concentration LCAT was inhibited. Small liposomes with phosphatidylcholine/cholesterol molar ratios of 3:1 up to 8.4:1 did not show any significant differences in the LCAT reaction, when used as substrates in the presence of various amounts of apoA-I and albumin. In contrast, the LCAT activity was significantly reduced by liposomes with phosphatidylcholine/cholesterol molar ratios below 3:1.
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D000818:Animals; D000906:Antibodies; D001053:Apolipoproteins / Q000097:blood; D004789:Enzyme Activation; D006801:Humans; D005779:Immunodiffusion; D007074:Immunoglobulin G / Q000494:pharmacology; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000097:blood* / Q000276:immunology; D013379:Substrate Specificity; D013552:Swine
D000906:Antibodies; D001053:Apolipoproteins; D007074:Immunoglobulin G; D007862:Phosphatidylcholine-Sterol O-Acyltransferase