Purification of human plasma lecithin:cholesterol acyltransferase and its activation by metal ions.
Lecithin:cholesterol acyltransferase (LCAT) has been purified from human plasma by sequential preparative ultracentrifugation, ion-exchange chromatography on DEAE-Sephacel, and affinity chromatography on HDL-Sepharose and on wheat germ agglutinin-Sepharose. After the final step, which included preparative electrophoresis or alternatively, chromatography on hydroxylapatite, a purification of about 24 000-fold was obtained. The LCAT preparation was pure according to alkaline polyacrylamide and SDS-polyacrylamide gel electrophoresis and did not react against antisera to apo AI, AII, and D. The LCAT preparation obtained by preparative electrophoresis was stimulated by Cu2+, Ni2+, Co2+, and Zn2+ at both stages of the reaction, phospholipase reaction and cholesterol esterification. This stimulatory effect was abolished by EDTA.
D002118:Calcium / Q000494:pharmacology; D002413:Cations, Divalent / Q000494:pharmacology*; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003035:Cobalt / Q000494:pharmacology; D003300:Copper / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006801:Humans; D009532:Nickel / Q000494:pharmacology; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000097:blood / Q000302:isolation & purification*; D015032:Zinc / Q000494:pharmacology
D002413:Cations, Divalent; D003035:Cobalt; D003300:Copper; D009532:Nickel; D007862:Phosphatidylcholine-Sterol O-Acyltransferase; D015032:Zinc; D002118:Calcium