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Fig. 6. The RGD motif is involved in DAB2-mediated inhibition of agonist-induced platelet aggregation. (A) DAB2 selectively inhibits agonist-induced platelet aggregation. Platelet aggregation was induced by the indicated concentration of agonists in the presence of GST, DAB2-PTB or DAB2-D66E (100 µg/ml). The platelet aggregation curves (left panel) and the percent of aggregation (right panel) were obtained and quantified by using a platelet aggregometer. (B) Schematic representation of the putative thrombin-cleavage sites in human DAB2-PTB and rat B. The putative thrombin-cleavage sites and the size of the cleavage product for human DAB2-PTB and rat B were shown. (C) Thrombin is a DAB2 protease. The recombinant human DAB2-PTB and in vitro transcription- and translation-derived [S³⁵]-methionine-labeled rat B were incubated in the absence (C) or presence (T) of thrombin (10 U/ml). Aliquots of the reaction mixtures were fractionated on SDS-PAGE and were visualized by Commassie Blue staining (DAB2-PTB) or autoradiography (B). (D) The DAB2-RGD peptide inhibits agonist-induced platelet aggregation. Platelet aggregation was induced by agonists at indicated concentration in the presence of DAB2-RGD or DAB2-RGE peptides (100 µg/ml). The platelet aggregation curves (left panel) and the percent of aggregation (right panel) was obtained and quantified by using a platelet aggregometer. The data represent the mean ± s.d. of three to four experiments. *P<0.05 and **P<0.001, compared with platelet aggregation induced by the same agonist in the presence of control GST protein (A) or DAB2-RGE (D). NS, not significant.

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