Transcytosis of lipoprotein lipase across cultured endothelial cells requires both heparan sulfate proteoglycans and the very low density lipoprotein receptor.
Lipoprotein lipase (LPL), the major enzyme responsible for the hydrolysis of circulating lipoprotein triglyceride molecules, is synthesized in myocytes and adipocytes but functions while bound to heparan sulfate proteoglycans (HSPGs) on the luminal surface of vascular endothelial cells. This requires transfer of LPL from the abluminal side to the luminal side of endothelial cells. Studies were performed to investigate the mechanisms of LPL transcytosis using cultured monolayers of bovine aortic endothelial cells. We tested whether HSPGs and members of the low density lipoprotein (LDL) receptor superfamily were involved in transfer of LPL from the basolateral to the apical side of cultured endothelial cells. Heparinase/heparinitase treatment of the basolateral cell surface or addition of heparin to the basolateral medium decreased the movement of LPL. This suggested a requirement for HSPGs. To assess the role of receptors, we used either receptor-associated protein, the 39-kDa inhibitor of ligand binding to the LDL receptor-related protein and the very low density lipoprotein (VLDL) receptor, or specific receptor antibodies. Receptor-associated protein reduced (125)I-LPL and LPL activity transfer across the monolayers. When the basolateral surface of the cells was treated with antibodies, only anti-VLDL receptor antibodies inhibited transcytosis. Moreover, overexpression of the VLDL receptor using adenoviral-mediated gene transfer increased LPL transcytosis. Thus, movement of active LPL across endothelial cells involves both HSPGs and VLDL receptor.
Obunike
J C
JC
Department of Medicine, Division of Preventive Medicine and Nutrition, Columbia University College of Physicians and Surgeons, New York, New York 10032, USA.
Lutz
E P
EP
Li
Z
Z
Paka
L
L
Katopodis
T
T
Strickland
D K
DK
Kozarsky
K F
KF
Pillarisetti
S
S
Goldberg
I J
IJ
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology*; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D026541:Heymann Nephritis Antigenic Complex; D006358:Hot Temperature; D007457:Iodine Radioisotopes; D008071:Lipoprotein Lipase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism; D021381:Protein Transport; D011973:Receptors, LDL / Q000378:metabolism*
D019812:Heparan Sulfate Proteoglycans; D026541:Heymann Nephritis Antigenic Complex; D007457:Iodine Radioisotopes; D008562:Membrane Glycoproteins; D011973:Receptors, LDL; C036053:VLDL receptor; D008071:Lipoprotein Lipase