Human carcinoma cells express receptors for distinct domains of thrombospondin.
Thrombospondin (TSP), an adhesive glycoprotein, is incorporated into the extracellular matrix, mediates cell attachment and spreading, chemotaxis, haptotaxis, and may participate in the directed movement of cells in metastasis. Evidence from several model systems suggests that these functions may be mediated by different domains within the TSP molecule. Radioligand binding assays on 11B squamous carcinoma cells with 125I-Radioligand binding assays on 11B squamous carcinoma cells with 125I-TSP demonstrated the presence of 1.2 x 10(6) sites/cell with an apparent Kd of 74 nM. Binding studies using TSP fragments demonstrated that both the NH2 terminal heparin-binding domain (HBD) and the COOH terminal fragment with a molecular weight of 140,000 (140K) retained the ability to bind 11B cells in a time-dependent, dose-dependent, saturable, and specific manner. The HBD bound to 11B cells with an apparent Kd of 1.2 microM at 1.4 x 10(6) sites/cell. Binding of 140K to cells demonstrated half-maximal binding at 36 nM and a Bmax of 1.9 x 10(5) sites/cell. The binding of 140K also showed a high degree of positive cooperativity with a Hill slope of +3.5, suggesting that binding one 140K molecule to cells leads to increased binding of additional 140K molecules. In addition, the HBD and 140K showed no cross-competition in binding assays. Therefore, it appears likely that these distinct TSP domains bind to separate sites on the cell surface. Neither vitronectin or the peptide RGDS were able to inhibit the binding of TSP or 140K to 11B cells. Based on these data, there appears to be more than one distinct receptor on 11B cells for TSP; one receptor class which mediates the binding of the HBD and a second receptor class which mediates the binding of the Mr 140,000 fragment.
Yabkowitz
R
R
Department of Pathology, University of Michigan Medical School, Ann Arbor 48109.
Dixit
V M
VM
D001665:Binding Sites; D018955:CD36 Antigens; D002294:Carcinoma, Squamous Cell / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D006493:Heparin / Q000378:metabolism; D006801:Humans; D009842:Oligopeptides / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism* / Q000494:pharmacology; D016024:Receptors, Cytoadhesin / Q000032:analysis*; D019699:Thrombospondins; D014407:Tumor Cells, Cultured
D018955:CD36 Antigens; D009842:Oligopeptides; D010980:Platelet Membrane Glycoproteins; D016024:Receptors, Cytoadhesin; D019699:Thrombospondins; C047981:arginyl-glycyl-aspartic acid; D006493:Heparin