Cellular localization and interaction of ABCA1 and caveolin-1 in aortic endothelial cells after HDL incubation

The goal of this study was to investigate the cellular localization and the interaction between caveolin-1 and ABCA1 in cholesterol-loaded aortic endothelial cells after HDL incubation. Immunofluorescence confocal microscopy showed that ABCA1 was found primarily on the cell surface, whereas caveolin-1 was revealed on the cell surface and in the cytoplasm. The HDL appeared to colocalize with ABCA1 and caveolin-1 on the cell surface. No free HDL was revealed in the cytoplasm. The HDL was colocalized neither with early endosome marker (CD71) nor with late endosome marker (LAMP2). The chemical cross-linking and immunoprecipitation analysis revealed that ABCA1 binds directly to both HDL and caveolin-1, whereas HDL does not bind directly to caveolin-1. The studies provide evidence for a direct interaction between ABCA1 and HDL, ABCA1 and caveolin-1, but not HDL and caveolin-1, indicating that ABCA1 may act as a structural platform between HDL and caveolin-1 on the cell surface during cellular cholesterol efflux.