Vascular endothelial cells undergo albumin endocytosis using a set of albumin binding proteins. This process is important for maintaining cellular homeostasis. We showed by several criteria that the previously described 73-kDa endothelial cell surface albumin binding protein is the 75-kDa transforming growth factor (TGF)-β receptor type II (TβRII). Albumin coimmunoprecipitated with TβRII from a membrane fraction from rat lung microvascular endothelial cells. Albumin endocytosis-negative COS-7 cells became albumin endocytosis competent when transfected with wild-type TβRII but not when transfected with a domain-negative kinase mutant of TβRII. An antibody specific for TβRII inhibited albumin endocytosis. A mink lung epithelial cell line, which expresses both the TGF-β receptor type I (TβRI) and the TβRII receptor, exhibited albumin binding to the cell surface and endocytosis. In contrast, mutant L-17 and DR-26 cells lacking TβRI or TβRII, respectively, each showed a dramatic reduction in binding and endocytosis. Albumin endocytosis induced Smad2 phosphorylation and Smad4 translocation as well as increased protein expression of the inhibitory Smad, Smad7. We identified regions of significant homology between amino acid sequences of albumin and TGF-β, suggesting a structural basis for the interaction of albumin with the TGF-β receptors and subsequent activation of TβRII signaling. The observed albumin-induced internalization of TβRII signaling may be an important mechanism in the vessel wall for controlling TGF-β responses in endothelial cells.