Leach Katie, Sexton Patrick M, Christopoulos Arthur
Drug Discovery Biology, Monash Institute of Pharmaceutical Science and Department of Pharmacology, Monash University, Parkville, Victoria, Australia.
Curr Protoc Pharmacol. 2011 Mar;Chapter 1:Unit 1.22. doi: 10.1002/0471141755.ph0122s52.
Allosteric interactions involve the simultaneous binding of two ligands to the same receptor. An allosteric modulator causes a conformational change in the receptor protein that yields a change in the binding or signaling of an orthosteric agent, i.e., an agonist or competitive antagonist that binds to the endogenous agonist binding site. Because of the complex nature of allosteric phenomena, the detection and quantification of their effects on orthosteric ligand binding relies on the use of both equilibrium and non-equilibrium assays to ensure proper interpretation of the findings. Outlined in this unit are the most common experimental approaches for measuring allosteric effects on orthosteric ligand affinity at G protein-coupled receptors. There is also a discussion of the analysis of experimental data derived from such assays.
变构相互作用涉及两个配体同时结合到同一受体上。变构调节剂会导致受体蛋白发生构象变化，从而使正构剂（即结合到内源性激动剂结合位点的激动剂或竞争性拮抗剂）的结合或信号传导发生改变。由于变构现象的复杂性，检测和量化它们对正构配体结合的影响依赖于使用平衡和非平衡测定法，以确保对研究结果的正确解释。本单元概述了用于测量G蛋白偶联受体上变构对正构配体亲和力影响的最常见实验方法。还讨论了对此类测定获得的实验数据的分析。
