title: Effect of substrate properties on the activity of lysosomal cholesteryl ester hydrolase.
abstract: The effects of the substrate properties on the catalytic activity of lysosomal cholesteryl ester hydrolase from rat liver have been examined with three standard substrate types: vesicle, micelle and emulsion. The pH optimum of the enzyme coincided to 4.5--5.0 with the substrate types employed. The apparent Km values were 15.3, 14.3 and 7.3 microM for vesicle, micelle and emulsion substrates, respectively. In the systems used in this study reaction products, cholesterol and oleic acid, and the nonionic surfactant Tween 80 and Triton X-100 Had an inhibitory effect. The emulsifier phosphatidylcholine and the charged phospholipid phosphatidic acid stimulated the activity. The mixed micelle of sodium taurocholate and phosphatidylcholine was the most potent substrate vehicle. With dipalmitoyl phosphatidylcholine vesicles the enzyme showed maximal activity at the gel-liquid-crystalline transition temperature of the phospholipid. The possible physiological significance of the lysosomal cholesteryl ester hydrolase is discussed with special reference to the form of the substrate.
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D002788:Cholesterol Esters / Q000378:metabolism; D004655:Emulsions; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology*; D008823:Micelles; D009829:Oleic Acids / Q000378:metabolism; D010712:Phosphatidic Acids / Q000494:pharmacology; D010743:Phospholipids / Q000494:pharmacology; D051381:Rats; D002787:Sterol Esterase / Q000378:metabolism*; D013501:Surface-Active Agents / Q000494:pharmacology; D013696:Temperature
chemical_list: D002788:Cholesterol Esters; D004655:Emulsions; D008823:Micelles; D009829:Oleic Acids; D010712:Phosphatidic Acids; D010743:Phospholipids; D013501:Surface-Active Agents; D002265:Carboxylic Ester Hydrolases; D002787:Sterol Esterase; D002118:Calcium