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- W2090900805 abstract "The use of the radioligand [35S]adenosine 5'-O-(2-thiodiphosphate) (ADPβ35S) for the determination of P2y-purinoceptors on turkey erythrocyte membranes has recently been described. In the present study, we were able to demonstrate specific binding of this radioligand in intact rat liver parenchymal cells. Within 10 min a thermodynamic equilibrium was obtained which lasted for 25 min with a subsequent decline. Displacement studies with several nucleotides were performed yielding Ki values of 1.5 ± 0.47 μM for UTP, 1.8 ± 0.35 μM for adenosine 5'-O-(2-thiodiphosphate) (ADPβS), 31 ± 6.2 μM for ATP and 35 ± 6.1 μM for GTP. In addition, we showed that ADPβ35S is highly resistant to degradation by ecto-nucleotidases, with only 14.5 ± 1.4% of total ADPβ35S present being degraded after 1 hr, and that the binding of ADPβ35S to its binding sites was modulated by EDTA. The Ki value of ATP shifted to 8.1 ± 1.2 μM upon the addition of 1 mM EDTA to the incubation medium. In these rat liver parenchymal cells all nucleotides promoted calcium entry in a dose-dependent manner with ec50 values of 3.5 ± 0.22 μM for UTP, 20.7 ± 3.1 μM for ATP, 38.3 ± 6.4 jμM for ADPβS and 73.6 ± 13.7 μM for GTP, with GTP being a partial agonist. Based on the data derived from the present study we discuss the possible correlation between binding and functional experiments and conclude that the described receptor resembles most closely the P2u-purinoceptor and/or “nucleotide receptor”, in that UTP is at least as active as ATP." @default.
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- W2090900805 date "1993-02-01" @default.
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- W2090900805 title "Binding of the radioligand [3Sadenosine 5'-O-(2-thiodiphosphate) and intracellular calcium response in rat liver parenchymal cells" @default.
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- W2090900805 doi "https://doi.org/10.1016/0006-2952(93)90162-p" @default.
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