创新的功能性 cAMP 检测法用于研究 G 蛋白偶联受体：在 GPR17 的药理学特征研究中的应用。
Innovative functional cAMP assay for studying G protein-coupled receptors: application to the pharmacological characterization of GPR17.
机构信息
Medicinal Chemistry Unit, School of Pharmacy, University of Camerino, 62032, Camerino, Italy.
出版信息
Purinergic Signal. 2011 Dec;7(4):463-8. doi: 10.1007/s11302-011-9245-8. Epub 2011 Jul 20.
Abstract
In this work, an innovative and non-radioactive functional cAMP assay was validated at the GPR17 receptor. This assay provides a simple and powerful new system to monitor G protein-coupled receptor activity through change in the intracellular cAMP concentration by using a mutant form of Photinus pyralis luciferase into which a cAMP-binding protein moiety has been inserted. Results, expressed as EC(50) or IC(50) values for agonists and antagonists, respectively, showed a strong correlation with those obtained with [(35)S]GTPγS binding assay, thus confirming the validity of this approach in the study of new ligands for GPR17. Moreover, this method allowed confirming that GPR17 is coupled with a G(αi).
摘要
在这项工作中，在 GPR17 受体上验证了一种创新的非放射性功能性 cAMP 测定法。该测定法通过使用插入了 cAMP 结合蛋白部分的 Photinus pyralis 荧光素酶的突变形式，提供了一种简单而强大的新系统，通过细胞内 cAMP 浓度的变化来监测 G 蛋白偶联受体活性。结果以激动剂和拮抗剂的 EC50 或 IC50 值表示，与用 [(35)S]GTPγS 结合测定法获得的值具有很强的相关性，从而证实了该方法在研究 GPR17 的新型配体中的有效性。此外，该方法还证实 GPR17 与 G(αi)偶联。
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