G蛋白偶联受体高阶聚化和信号转导中蛋白复合体的时空动态检测进展

Spatio-temporal dynamic detection of higher-order G protein-coupled receptor oligomers and protein complex in signalling

传统观点认为,G蛋白偶联受体与G蛋白、调节蛋白、GRKs以及arrestins间的相互作用是连续的,其中涉及多个蛋白质之间快速的集合和解聚。而实际上,GPCRs和相关靶分子之间的作用是通过一系列细胞内蛋白质-蛋白质相互作用的信号转导网络来实现的。近10年,随着一些新兴技术的出现,如FRET、BRET和BiFC,实时动态观测二个蛋白质间相互作用的研究成为热点。最近,一些技术结合的方法为从"时间、空间、动态、连续"检测更多蛋白质之间的相互作用的研究拉开新的帷幕。从横向来看,这些技术的结合可以用来研究细胞膜上3种甚至更多蛋白质的高阶寡聚化;从纵向来看,可以对细胞膜-细胞质-细胞核中所发生的信号转导网络的研究提供新的技术手段,多种技术结合所发挥作用是最近出现的几种技术所无法企及的,这对于疾病发病机制的研究及新的药物靶点的发现具有深远意义。因此,本文就一些技术的结合及其应用做一简要综述。

Traditionally,G-protein-coupled receptor（GPCR） interactions with their G proteins and regulatory proteins,GPCR kinases（GRKs） and arrestins,are described as sequential events involving rapid assemblies/disassemblies.But in fact,the events leading to the interaction of GPCRs with targeted molecules are generally believed to follow a sequential scheme.In the past decade,with the emergence of novel technology,especially FRET and BRET,real-time dynamic observation of the interaction between two proteins became hot.Rencently,detection of interaction among proteins was drawn open by some combined technologies from the ＂time,space,dynamic and continuous＂.From the horizontal point of view,these combined technologies can be used to study the oligomers formed by three or more proteins in membrane;from the vertical perspective,it provides novel techniques for the network of signal transduction in membrane-cytoplasm-nucleus.Combined technologies play an important role while several latest techniques do not,which is far-reaching for disease pathogenesis and the emergence of new drug targets.Consequently,we review some combined techniques as well as their applications.