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Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA 19104
1 To whom correspondence should be addressed. e-mail: glickj{at}mail.med.upenn.edu
Endothelial lipase (EL) is a new member of the triglyceride lipase gene family previously reported to have phospholipase activity. Using radiolabeled lipid substrates, we characterized the lipolytic activity of this enzyme in comparison to lipoprotein lipase (LPL) and hepatic lipase (HL) using conditioned medium from cells infected with recombinant adenoviruses encoding each of the enzymes. In the absence of serum, EL had clearly detectable triglyceride lipase activity. Both the triglyceride lipase and phospholipase activities of EL were inhibited in a dose-dependent fashion by the addition of serum. The ratio of triglyceride lipase to phospholipase activity of EL was 0.65, compared with ratios of 24.1 for HL and 139.9 for LPL, placing EL at the opposite end of the lipolytic spectrum from LPL. Neither lipase activity of EL was influenced by the addition of apolipoprotein C-II (apoC-II), indicating that EL, like HL, does not require apoC-II for activation. Like LPL but not HL, both lipase activities of EL were inhibited by 1 M NaCl. The relative ability of EL, versus HL and LPL, to hydrolyze lipids in isolated lipoprotein fractions was also examined using generation of FFAs as an end point.
As expected, based on the relative triglyceride lipase activities of the three enzymes, the triglyceride-rich lipoproteins, chylomicrons, VLDL, and IDL, were efficiently hydrolyzed by LPL and HL. EL hydrolyzed HDL more efficiently than the other lipoprotein fractions, and LDL was a poor substrate for all of the enzymes.
Abbreviations: AdGFP, adenovirus encoding GFP; AdhEL, adenovirus encoding human endothelial lipase; AdhHL, adenovirus encoding human hepatic lipase; AdhLPL, adenovirus encoding human LPL; DPPC, dipalmitoylphosphatidylcholine; chylo, chylomicrons; EL, endothelial lipase; EC, esterified cholesterol; FC, free cholesterol; GFP, green fluorescent protein; HL, hepatic lipase; LPPC, lysopalmitoylphosphatidylcholine; PL, phospholipid; PLA2, phospholipase A2
Supplementary key words lipoprotein lipase • hepatic lipase • triglyceride lipase family • phospholipase • apoC-II • heparin
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