SECRETION-CAPTURE ROLE FOR APOLIPOPROTEIN-E IN REMNANT LIPOPROTEIN METABOLISM INVOLVING CELL-SURFACE HEPARAN-SULFATE PROTEOGLYCANS

To determine the impact of enhanced apolipoprotein (apo) E secretion o n the mechanism of remnant lipoprotein uptake, rat hepatoma cells (McA -RH7777) were stably transfected with normal human apoE3 or receptor b inding-defective apoE-Leiden. After a 2-h incubation, the human apoE s ecreted from the transfected hepatocytes was 10-12 times greater than the endogenous rat apoE. The apoE3-transfected cells bound and interna lized rabbit beta-very low density lipoproteins (beta-VLDL) to a much greater degree than did apoE-Leiden-transfected cells and nontransfect ed cells. The apoE3-secreting cells displayed a 2-3.5-fold enhancement of cell-associated beta-VLDL compared to either the apoE-Leiden-trans fected or nontransfected cells. Fluorescently labeled beta-VLDL were o bserved to concentrate within intracellular granules of the apoE3-tran sfected cells, presumably within endosomes and lysosomes. Furthermore, electron microscopy revealed that the apoE3-secreting cells displayed abundant beta-VLDL and chylomicron remnants on their cell surfaces an d microvilli, in contrast to non-transfected or apoE-Leiden-secreting cells. Electron microscopy also revealed an abundance of chylomicron r emnants within intracellular vesicles and multivesicular bodies of apo E3-transfected hepatocytes. Heparinase treatment (3 units/ml) complete ly abolished the increased association of beta-VLDL with apoE3-transfe cted cells but did not affect the limited association of beta-VLDL wit h apoE-Leiden-transfected or nontransfected cells. We established that the apoE3-enriched beta-VLDL were bound to cell surface heparan sulfa te proteoglycans, as was the newly synthesized and secreted apoE3 (app roximately 12% of the total secreted apoE3 was released by heparinase and suramin; 4% by heparin). In addition, reisolation of beta-VLDL by fast performance liquid chromatography after their incubation with exo genous apoE3, with medium from apoE3-secreting cells, or with the apoE 3-secreting cells themselves revealed that the particles were enriched in apoE3 and displayed enhanced binding. These results suggest a secr etion-capture role for apoE and indicate an important role for heparan sulfate proteoglycans on the cell surface for remnant lipoprotein met abolism.