INVESTIGATION OF PLASMA MEMBRANE-ASSOCIATED APOLIPOPROTEIN-E IN PRIMARY MACROPHAGES

Our previous studies identified the lysosome as the compartment for de gradation of newly synthesized apoE in primary macrophages. Lysosomal degradation of newly synthesized apoE is extensive and rapid (>50% in 60 min). In the present study we tested the hypothesis that the macrop hage cell surface is part of the itinerary of apoE in its path to the lysosomes. We therefore examined the existence and size of the apoE po ol associated with the macrophage cell surface. Such a pool may not on ly provide a mechanism of targeting apoE for lysosomal degradation, by endocytosis, but also have important implications for the metabolism of lipoproteins by macrophages. Treatment of macrophages with heparin (10 mu g/ml and 5 mg/ml) and heparinase I (1 U/ml), which releases sub stantial amounts of apoE from HepG2 cells, results in no additional re lease of apoE from macrophages. Treatment of macrophages with xyloside (1 mM) or GRGDTP (500 mu g/ml) does not decrease the extent of cell-a ssociated apoE. Both immunogold labeling, followed by electron microsc opy, and immunofluorescent labeling and light microscopy further confi rm the lack of significant amounts of cell surface-associated apoE in macrophages. In contrast, immunolabeled apoE is readily observed in pe rmeabilized cells. Taken together, these data indicate the absence of significant apoE-glycosaminoglycan interaction at the macrophage cell surface. The lack of such an interaction is likely due to a paucity of heparan sulfate proteoglycans on the macrophage cell surface, when co mpared to hepatocytes. Along with our previous observations (Deng, J., V. Rudick, and L. Dory, 1995. J. Lipid Res. 36: 2129-2140), these res ults suggest direct targeting of a portion of newly synthesized apoE f rom trans-Golgi network to lysosomes for degradation, without involvin g the plasma membrane and endocytosis.