Characterization of the platelet agglutinating activity of thrombospondin.
Thrombospondin (TSP) is a glycoprotein secreted from the alpha-granules of platelets upon activation. In the presence of divalent cations, the secreted protein binds to the surface of the activated platelets and is responsible for the endogenous lectin-like activity associated with activated platelets. Platelets fixed with formaldehyde following activation by thrombin are agglutinated by exogenously added TSP. Fixed, nonactivated platelets are not agglutinated. The platelet agglutinating activity of TSP is optimally expressed in the presence of 2 mM each of Mg2+ and Ca2+. Reduction of the disulfide bonds within the TSP molecule inhibits its platelet agglutinating activity. TSP bound to the surface of fixed, activated platelets can be eluted by the addition of disodium ethylenediaminetetraacetate. This approach was exploited to identify the region of the TSP molecule containing the platelet binding site. The binding site resides within a thermolytic fragment of TSP with Mr 140 000 but is not present in the Mr 120 000 fragment derived from the polypeptide of Mr 140 000. Since both the Mr 140 000 and 120 000 fragments contain fibrinogen binding sites, this finding suggests that the binding of TSP to the platelet surface requires interaction with other platelet surface components in addition to fibrinogen. The observation that fibrinogen only partially inhibits the TSP-mediated agglutination of fixed, activated platelets is consistent with this interpretation.
D001792:Blood Platelets / Q000502:physiology*; D002118:Calcium / Q000494:pharmacology; D002413:Cations, Divalent; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D010446:Peptide Fragments / Q000302:isolation & purification / Q000502:physiology; D010974:Platelet Aggregation / Q000187:drug effects; D013820:Thermolysin; D019699:Thrombospondins
D002413:Cations, Divalent; D006023:Glycoproteins; D010446:Peptide Fragments; D019699:Thrombospondins; D013820:Thermolysin; D008274:Magnesium; D002118:Calcium