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G蛋白偶联受体(G-protein-coupled receptors,GPCRs)是迄今发现的最大的多药物靶点的受体超家族,其激动剂或拮抗剂常被用于治疗各种疾病,在药物研发中扮演重要角色。测定配体与受体亲和力的结合实验是药物评价、研发和作用机理研究中不可缺少的部分。用于研究GPCRs与其配体结合的分析技术,可以按照标记方式的异同分为三类:同位素标记法、非同位素标记法和非标记法。非同位素标记法多采用荧光配体标记,利用竞争法或荧光共振能量转移直接获知结合情况;非标记法中利用各种生物传感器,通过检测配体结合受体后胞内分子运动引起的电信号或光信号的变化来表征结合情况。此外,质谱也已被用于受体-配体平衡解离常数的测定,显示了其在非标记结合分析及受体功能研究中的潜力。本文归纳了目前用于GPCRs结合分析的主要技术,对其原理及优缺点进行了综述,并对受体-配体结合分析中新方法的应用进行了展望。
G-protein-coupled receptors (GPCRs) are by far the largest multi-drug receptor superfamily discovered and their agonists or antagonists are often used in the treatment of various diseases. In drug development Play an important role. The binding assay to determine the affinity of a ligand to a receptor is an integral part of the drug evaluation, development, and mechanism of action. The analytical techniques used to study the binding of GPCRs to their ligands can be grouped into three categories based on the similarities and differences in labeling methods: isotope labeling, non-isotopic labeling and non-labeling. Non-isotope labeling method using fluorescent ligand tag, the use of competitive method or fluorescence resonance energy transfer directly informed of the binding; non-labeled method using a variety of biosensors, by detecting the ligand binding to the receptor after the intramolecular motility caused by electrical signals Or changes in optical signals to characterize the binding. In addition, mass spectrometry has also been used for the determination of receptor-ligand equilibrium dissociation constants, indicating its potential for non-labeled binding assays and receptor function studies. This article summarizes the main techniques currently used for binding GPCRs, summarizes the principles, advantages and disadvantages of GPCRs, and looks into the application of new methods in receptor-ligand binding assays.