Lipoprotein lipase- and phospholipase A2-catalyzed hydrolysis of phospholipid vesicles with an encapsulated fluorescent dye. Effects of apolipoproteins.
The self-quenching dye, 6-carboxyfluorescein, has been encapsulated into sonicated vesicles of egg phosphatidylcholine. Porcine pancreatic phospholipase A2 and bovine milk lipoprotein lipase catalyze the hydrolysis of the phosphatidylcholine resulting in the release of the encapsulated dye and a large increase in 6-carboxyfluorescein fluorescence. The fluorescence increase occurs in parallel with the formation of lysophosphatidylcholine and is strongly dependent on Ca2+ for phospholipase A2 catalysis and on apolipoprotein C-II for hydrolysis by lipoprotein lipase. Other apolipoproteins, including apolipoproteins C-III, C-I, and A-I, do not enhance lipoprotein lipase activity towards this substrate. We conclude that the enhancement of lipoprotein lipase activity by apolipoprotein C-II is a specific property of the activator protein due to its interaction with lipoprotein lipase or an enzyme/lipid interface and not a characteristic of lipid-binding proteins in general.
D000818:Animals; D001056:Apolipoproteins C / Q000097:blood*; D002417:Cattle; D005260:Female; D005452:Fluoresceins; D006801:Humans; D007700:Kinetics; D008071:Lipoprotein Lipase / Q000378:metabolism*; D008081:Liposomes; D008892:Milk / Q000201:enzymology; D010713:Phosphatidylcholines; D010740:Phospholipases / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D011663:Pulmonary Surfactants; D013050:Spectrometry, Fluorescence
D001056:Apolipoproteins C; D005452:Fluoresceins; D008081:Liposomes; D010713:Phosphatidylcholines; D011663:Pulmonary Surfactants; C024098:6-carboxyfluorescein; D010740:Phospholipases; D010741:Phospholipases A; D008071:Lipoprotein Lipase; D054467:Phospholipases A2