Incorporation of hormone-sensitive lipase into phosphatidylcholine vesicles.
Enzymatically active, detergent-solubilized purified hormone-sensitive lipase (HSL) was incorporated into phosphatidylcholine (PC) vesicles, using a detergent-dialysis procedure with small PC vesicles, obtained by sonication, as phospholipid source and CHAPS, a zwitterionic bile-salt derivative, as detergent. Association of enzyme protein with the PC vesicles was verified by floatation in a discontinuous dextran gradient and by gel chromatography. An average of 35% of added HSL was incorporated into the vesicles. The vesicles were shown, by quasi-elastic light scattering and electron microscopy, to have a diameter of approximately 160 nm. The vesicle-associated HSL could be phosphorylated by cyclic AMP-dependent protein kinase. The vesicles were stable, both with regard to enzyme activity and size, for at least 4 days when stored at 4 degrees C. The preparation of detergent-free, vesicle-associated and stable HSL provides new possibilities to study some of its properties, and supports and extends the previous report (Holm, C., Fredrikson, G., and Belfrage, P., J. Biol. Chem. 261, 15659-15661, 1986) which demonstrated the amphiphilic character of HSL.
Department of Medical and Physiological Chemistry, University of Lund, Sweden.
C
C
Holm
G
G
Fredrikson
R
R
Sundler
P
P
Belfrage
D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D002793:Cholic Acids / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology; D003911:Dextrans; D003956:Dialysis / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D008027:Light; D008049:Lipase / Q000378:metabolism*; D008567:Membranes, Artificial; D008854:Microscopy, Electron; D010713:Phosphatidylcholines / Q000378:metabolism*; D051381:Rats; D012542:Scattering, Radiation; D002787:Sterol Esterase / Q000378:metabolism*
D002793:Cholic Acids; D003902:Detergents; D003911:Dextrans; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D002265:Carboxylic Ester Hydrolases; D002787:Sterol Esterase; D008049:Lipase; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate