Dynamic and stable pools of apoE differ functionally at the HepG2 cell surface.
Synthesis of apolipoprotein (apo)E in hepatocytes leads to both secretion and retention in cell surface pools. Inclusion of Brefeldin A to HepG2 cells prompted a rapid decrease of cell surface apoE to about 37% of control values after a 3-h incubation. The t(1/2) for this dynamic pool was estimated to be 15 min. In contrast, a stable fraction of apoE (t(1/2) > 20 h) was found in association with the extracellular matrix (ECM). Increased content of apoE on the ECM correlated with decreased binding of VLDL. Decreased apoE on the cell surface correlated with increased binding of VLDL to cells. Collectively, this suggests that glycosaminoglycan-bound apoE can occlude binding sites for apoE-containing lipoproteins on glycosaminoglycans. In solid-phase assays, heparin, suramin, and chondroitin sulfates A and B efficiently inhibited the binding of apoE to heparan sulfate proteoglycans, but were unable to displace apoE from this glycosaminoglycan. Finally, decreasing cell surface apoE with suramin subsequently decreased the apoE content on secreted apoB-containing lipoproteins without affecting the overall secretion of apoE or apoB to the extracellular medium. In summary, cell surface apoE comprises both dynamic fractions, which can be donated to newly secreted lipoproteins, and stable fractions, which may act to minimize the unproductive binding of lipoproteins to the ECM.
Burgess
J W
JW
Lipoprotein and Atherosclerosis Group, University of Ottawa Heart Institute, 40 Ruskin Street, Ottawa, Ontario, Canada K1Y 4W7.
Marcel
Y L
YL
D001057:Apolipoproteins E / Q000032:analysis / Q000378:metabolism*; D001665:Binding Sites; D020126:Brefeldin A / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000737:chemistry*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002809:Chondroitin Sulfates / Q000494:pharmacology; D003871:Dermatan Sulfate / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism; D006207:Half-Life; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008074:Lipoproteins / Q000378:metabolism; D008079:Lipoproteins, VLDL / Q000378:metabolism; D008113:Liver Neoplasms / Q000737:chemistry*; D013498:Suramin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D001057:Apolipoproteins E; D006025:Glycosaminoglycans; D008074:Lipoproteins; D008079:Lipoproteins, VLDL; D020126:Brefeldin A; D003871:Dermatan Sulfate; D013498:Suramin; D006493:Heparin; D002809:Chondroitin Sulfates