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Vol 50(2016) N 1 p. 111-117; DOI 10.1134/S0026893316010180 O.V. Sirotkina1,2,3,4*, A.B. Laskovets4, P.A. Andoskin1,3, A.K. Emelyanov1,3, A.M. Zabotina1, T.V. Vavilova2,4 Increase in GP IIb-IIIa and P2Y12 Receptors in Activated Platelets as the Possible Indicator of de novo Protein Synthesis 1Konstantinov St. Petersburg Institute of Nuclear Physics, Kurchatov Institute Scientific Research Center, Gatchina, Leningrad oblast, 188300 Russia2Almazov North-West Federal Medical Research Center, St. Petersburg, 197341 Russia 3Pavlov First St. Petersburg State Medical University, St. Petersburg, 197022 Russia 4Mechnikov North-West State Medical University, St. Petersburg, 191015 Russia *olga_sirotkina@mail.ru Received - 2015-07-09; Accepted - 2015-07-16 Although platelets lack nuclei, they are capable of de novo protein synthesis. We speculate that key platelet receptors are involved in the regulation of this process, and the changes in their number indicate the de novo protein synthesis in platelets. The object of our study was native platelets obtained from healthy donors. Using flow cytometry and Western blot, we determined the number of GP IIb-IIIa receptors (fibrinogen receptor) and P2Y12 receptors (ADP receptor) on the surface of platelets upon their activation with ADP and collagen. To verify the approaches and techniques used, we studied IL-1β protein, which was previously shown to be synthesized de novo in activated platelets. GP IIb-IIIa receptor numbers correlate with the number of P2Y12 receptors on the cell surface (R = 0.45, p = 0.03). It was demonstrated that the platelet receptor numbers are higher on the surface of the cells with high functional activity. According to the data obtained by Western blot, upon the cell activation with ADP, the number of GP IIb-IIIa and P2Y12 receptors increases, which may serve as evidence of these proteins being synthesized in the activated platelets. It was observed that the level of P2Y12 and IL-1β was lower in the samples where GP IIb-IIIa receptor was blocked by the selective inhibitor, i.e., the Fab fragment of the antibodies that specifically recognizes the GP IIb-IIIa complex. This suggests the important role of GP IIb-IIIa receptor in the regulation of protein synthesis. de novo protein synthesis, platelets, in vitro platelet activation, platelet receptor GP IIb-IIIa, platelet receptor P2Y12, flow cytometry, Western blott |