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首页> 外文期刊>Neurochemistry International: The International Journal for the Rapid Publication of Critical Reviews, Preliminary and Original Research Communications in Neurochemistry >Kinetic analysis of (35S)dATP alpha S interaction with P2y(1) nucleotide receptor.
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Kinetic analysis of (35S)dATP alpha S interaction with P2y(1) nucleotide receptor.

机译：（35S）dATP alpha S与P2y（1）核苷酸受体相互作用的动力学分析。

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摘要

The kinetics of 2'-deoxyadenosine-5'-O-(1-thiotriphosphate) ([(35)S]dATP alpha S) interaction with membrane fragments of transfected astrocytoma 1321N1 cells, expressing human P2Y(1) receptors, and the same wild-type cells, not expressing P2Y receptors were studied. Binding of this radioligand was observed with both types of membranes, but sites showing slow on-rate were found only on the transfected cells. These "slow" binding sites behaved as a kinetically homogeneous population and their interaction with the radioligand was shown to occur in two steps, R+A(K(A))<==>RA(k(i))<==>(k(-i))(RA), including the relatively slow isomerization of the complex RA into (RA). Evidence was obtained to assign the isomerized ("slow") binding sites on the transfected cells as P2Y(1) receptor sites, differentiated from other binding sites of non-receptor origin by kinetic analysis, and characterised by the kinetic parameters K(A)=59 +/- 19 nM, k(i)=(9.0 +/- 0.8)10(-3)s(-1) and k(-i)=(3.9 +/- 0.7)10(-3)s(-1). [(35)S]dATP alpha S binding, with kinetic criteria, can be of value for differentiation of the receptor sites from non-receptor sites and thus provides solid basis for radioligand assay of P2Y(1) receptors.
机译：2'-脱氧腺苷-5'-O-（1-硫代三磷酸）（[（35）S] dATP alpha S）与表达人P2Y（1）受体的转染星形细胞瘤1321N1细胞膜片段相互作用的动力学研究了不表达P2Y受体的野生型细胞。在两种类型的膜上都观察到了这种放射性配体的结合，但是仅在转染的细胞上发现了显示慢速开启的位点。这些“缓慢的”结合位点表现为动力学上均一的种群，它们与放射性配体的相互作用显示为两步发生，R + A（K（A））<==> RA（k（i））<==> （k（-i））（RA），包括将复合物RA相对缓慢地异构化为（RA）。获得的证据是将转染细胞上的异构化（“慢”）结合位点指定为P2Y（1）受体位点，通过动力学分析与非受体来源的其他结合位点区分开，并通过动力学参数K（A）进行了表征。 = 59 +/- 19 nM，k（i）=（9.0 +/- 0.8）10（-3）s（-1）和k（-i）=（3.9 +/- 0.7）10（-3）s （-1）。 [（35）S] dATPαS结合，具有动力学标准，对于区分受体位点与非受体位点可能具有价值，因此为P2Y（1）受体的放射性配体测定提供了坚实的基础。

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