Studying ligand efficacy at G protein-coupled receptors using FRET.
Studying ligand efficacy at G protein-coupled receptors using FRET.
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DOI：
10.1007/978-1-61779-160-4_6
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发表时间：
2011
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与G蛋白偶联受体(GPCRs)结合的药物“配体”可以通过稳定不同的受体构象，刺激、完全(完全激动剂)或部分(部分激动剂)或降低(反向激动剂)基础受体的活性。本征效应一词被用来表示配体激活其受体的能力，以及当不同的配体占据同一受体的相同部分时，区分不同配体的不同信号传递能力的参数。大多数方法使用下游的生化和生理反应作为“内在疗效”的替代，但不能在受体水平上直接测量它。在这里，我描述了Förster共振能量转移(FRET)方法的发展，该方法允许直接在GPCR水平上严格测量配体的内在有效性，并且独立于实验条件的变化。这种方法还允许将配体的内在效应与受体多态或受体异二聚化的影响联系起来。
Drug “ligands” that bind G protein-coupled receptors (GPCRs) can either stimulate, fully (full agonists) or partially (partial agonists), or reduce (inverse agonists) basal receptor activity, by stabilizing different receptor conformations. The term “intrinsic efficacy” was introduced as a parameter to express the ability of a ligand to activate its receptor and to differentiate the varying signaling capacity of diverse ligands when they occupy the same fraction of a single receptor. Most methods use downstream biochemical and physiological responses as proxies of “intrinsic efficacy” but cannot measure it directly at the level of the receptor. Here I describe the development of a Förster resonance energy transfer (FRET) approach that permits the rigorous measurement of the intrinsic efficacy of a ligand directly at the level of a GPCR and independent from variation in experimental conditions. This approach also allows intrinsic efficacies of ligands to be linked with the effects of receptor polymorphisms or receptor heterodimerization.