G蛋白偶联受体配体结合分析技术

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 so far the largest family of receptors of multi-drug targets,with their agonists and antagonists being used in the treatment of a wide range of diseases.Binding assay,in which the affinity between receptors and ligands is measured,is often a requisite in the research of GPCRs for both mechanism studies and drug development.The technologies used in the GPCR-ligand binding assays could be classified into three categories,namely:1)isotope labelled,2)non-isotope labelled and 3)label-free methods.In non-isotope labelled methods,fluorescent ligands are often used for direct measurement through competitive binding or fluorescence resonance energy transfer,while in label-free methods,biosensors are employed to measure the changes of electric or optical signals induced by the bindings between receptors and ligands.Recently mass spectrometry has also shown promising performance in binding assays.In this review,the mechanisms,accomplishments and future applications of these major technologies in GPCRs binding assays were summarized.