THE FUNCTIONAL THROMBIN RECEPTOR IS ASSOCIATED WITH THE PLASMALEMMA AND A LARGE ENDOSOMAL NETWORK IN CULTURED HUMAN UMBILICAL VEIN ENDOTHELIAL-CELLS

The functional thrombin receptor, normally expressed by endothelial ce lls and platelets, is a member of the G protein-coupled, seven membran e-spanning-domain receptor family and is thought to be responsible for most, if not all, the cell stimulatory effects of thrombin, Upon bind ing, thrombin cleaves the receptor's N-terminal ectodomain, unmasking a new N terminus, which by itself activates the receptor, Using antibo dies to different domains of the human thrombin receptor, we have loca lized the receptor in cultured human umbilical vein endothelial cells by indirect immunofluorescence and immunoelectron microscopy, We found the receptor expressed on the plasmalemma of cultured endothelial cel ls in individual units rather than in clusters, at lower concentration than, and at different sites from, thrombomodulin, We also found the receptor associated with a distinct, intracellular, transferrin recept or-containing, tubulovesicular network, The thrombin receptor-positive structure spread from the perinuclear region to the periphery of the cells, exhibiting a number of varicosities interconnected by branching tubular elements, strikingly similar to an image recently described f or a continuous endosomal reticulum, Our results provide morphological evidence for the presence of the functional thrombin receptor at rela tive low density on the surface of cultured endothelial cells (compare d to thrombomodulin) and in relatively large quantities inside the cel ls, associated with an endosomal compartment.