Co-clustering and internalization of low-density lipoproteins and alpha 2-macroglobulin in human skin fibroblasts

The endocytosis of low density lipoprotein (LDL) and α₂-macroglobulin (α₂M) has been examined simultaneously in human skin fibroblasts. Incubation of cells at 4 °C with rhodamine-α₂M and LDL plus [(dichlorotriazinyl)amino]fluorescein-anti-LDL gave a weak fluorescence for α₂M and a brighter, clustered fluorescence for LDL. Following warming to 37 °C, LDL and α₂M were observed to be coincident within endocytotic vesicles in the cell. By electron microscopy, LDL-ferritin and α₂M-colloidal gold were present in the same coated pit at 4 °C. After 7 min at 37 °C, both ligands were observed in the same receptosome. Pretreatment of fibroblasts at 37 °C with 200–300 μM dansylcadaverine or 50 mM methylamine blocked clustering and internalization of both LDL and α₂M. Bacitracin (5 mg ml^?) blocked clustering and endocytosis of α₂M, but not of LDL. These data indicate that both LDL and α₂M are processed via the same endocytotic pathway in skin fibroblasts.