Endogenously produced endothelial lipase enhances binding and cellular processing of plasma lipoproteins via heparan sulfate proteoglycan-mediated pathway.
Endothelial lipase (EL) is a new member of the triglyceride lipase gene family, which includes lipoprotein lipase (LpL) and hepatic lipase (HL). Enzymatic activity of EL has been studied before. Here we characterized the ability of EL to bridge lipoproteins to the cell surface. Expression of EL in wild-type Chinese hamster ovary (CHO)-K1 but not in heparan sulfate proteoglycan (HSPG)-deficient CHO-677 cells resulted in 3-4.4-fold increases of 125I-low density lipoprotein (LDL) and 125I-high density lipoprotein 3 binding (HDL3). Inhibition of proteoglycan sulfation by sodium chlorate or incubation of cells with labeled lipoproteins in the presence of heparin (100 microg/ml) abolished bridging effects of EL. An enzymatically inactive EL, EL-S149A, was equally effective in facilitating lipoprotein bridging as native EL. Processing of LDL and HDL differed notably after initial binding via EL to the cell surface. More than 90% of the surface-bound 125I-LDL was destined for internalization and degradation, whereas about 70% of the surface-bound 125I-HDL3 was released back into the medium. These differences were significantly attenuated after HDL clustering was promoted using antibody against apolipoprotein A-I. At equal protein concentration of added lipoproteins the ratio of HDL3 to VLDL bridging via EL was 0.092 compared with 0.174 via HL and 0.002 via LpL. In summary, EL mediates binding and uptake of plasma lipoproteins via a process that is independent of its enzymatic activity, requires cellular heparan sulfate proteoglycans, and is regulated by ligand clustering.
Fuki
Ilia V
IV
Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA. iliaf@mail.med.upenn.edu
Blanchard
Nadine
N
Jin
Weijun
W
Marchadier
Dawn H L
DH
Millar
John S
JS
Glick
Jane M
JM
Rader
Daniel J
DJ
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D008049:Lipase / Q000737:chemistry*; D008071:Lipoprotein Lipase / Q000378:metabolism; D008074:Lipoproteins / Q000097:blood* / Q000737:chemistry / Q000378:metabolism; D008075:Lipoproteins, HDL / Q000378:metabolism; D011485:Protein Binding; D013997:Time Factors
D019812:Heparan Sulfate Proteoglycans; D008024:Ligands; D008074:Lipoproteins; D008075:Lipoproteins, HDL; D008049:Lipase; D008071:Lipoprotein Lipase