Effects of detergents on the lecithin:cholesterol acyltransferase reaction.
This paper describes the effect of an ionic (sodium dodecyl sulfate; SDS) and a nonionic detergent (Triton X-100) on the substrate and enzyme components of the lecithin: cholesterol acyltransferase (LCAT) reaction. When the enzyme sources (purified or partially purified) or the respective substrates [high-density lipoproteins (HDL) or proteoliposomes] were preincubated with detergents, a consistent trend in LCAT activity was only seen when partially purified LCAT was used as the enzyme source. This trend indicated an approximately 25% increase in enzyme activity over the control when 10(-4) M SDS and 2 X 10(-3)% Triton X-100 were present in the preincubation mixtures, respectively. Those observations suggested that, during the preincubations and subsequent assays, the enzyme (in the presence of detergents) was allowed to dissociate from the endogenous substrate and subsequently interact with the exogenous substrate molecules. Additional experiments utilizing molecular-sieve chromatography with whole plasma and partially purified enzyme also showed that dissociation of LCAT/lipoprotein complexes occurred in the presence of detergent. SDS was also shown to enhance the reaction of LCAT in whole plasma with anti-LCAT antibody in an enzyme-linked immunoassay system, indicating that the detergent treatment facilitated the exposure of additional antigenic sites, perhaps via dissociation of the enzyme from plasma lipoproteins.
D000818:Animals; D016632:Apolipoprotein A-I; D001053:Apolipoproteins; D003902:Detergents / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D008075:Lipoproteins, HDL; D008081:Liposomes; D008970:Molecular Weight; D017830:Octoxynol; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011510:Proteolipids; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology*; D013501:Surface-Active Agents / Q000494:pharmacology*
D016632:Apolipoprotein A-I; D001053:Apolipoproteins; D003902:Detergents; D008075:Lipoproteins, HDL; D008081:Liposomes; D011510:Proteolipids; D013501:Surface-Active Agents; C020485:proteoliposomes; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D017830:Octoxynol; D007862:Phosphatidylcholine-Sterol O-Acyltransferase