Immunochemical identification of a thrombospondin-like structure in an arterial microfibrillar extract

Arterial microfibrils contain a 128 Kd collagenase and pepsin resistant glycoprotein (GP 128) essential for their ability to induce platelet aggregation. A previous report (Fauvel F. et al, (1984) Biochem. Biophys. Res. Comm., 123, 114-120) showed that GP 128 and thrombospondin (TSP) synthetized by endothelial cells each inhibited the aggregation of platelets by microfibrils and not by collagen. We used a monospecific antiplatelet TSP IgG in an immunoblotting assay for the identification of a TSP-like structure in untreated, collagenase-treated and pepsin-treated arterial microfibrils. The only constituent recognized in the three samples of microfibrils was GP 128. Fab fragments of this IgG provoked a dose dependent inhibition of the microfibril induced platelet aggregation (50% inhibition with 0.25 mg, 100% inhibition with 1 mg); in contrast, they did not affect collagen induced aggregation. The results indicate that a glycoprotein constituent with a thrombospondin-like antigenicity is involved in the thrombogenic properties of arterial microfibrils.