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Abstract：
G protein-coupled receptors (GPCRs) are membrane proteins that transmit specific external stimuli into cells by changing their conformation. This conformational change allows them to couple and activate G-proteins to initiate signal transduction. A critical challenge in studying and inferring these structural dynamics arises from the complexity of the cellular environment, including the presence of various endogenous factors. Due to the recent advances in cell-expression systems, membrane-protein purification techniques, and labeling approaches, it is now possible to study the structural dynamics of GPCRs at a single-molecule level both in vitro and in live cells. In this review, we discuss state-of-the-art techniques and strategies for expressing, purifying, and labeling GPCRs in the context of single-molecule research. We also highlight four recent studies that demonstrate the applications of single-molecule microscopy in revealing the dynamics of GPCRs. These techniques are also useful as complementary methods to verify the results obtained from other structural biology tools like cryo-electron microscopy and x-ray crystallography.
摘要：
G蛋白偶联受体（GPCRs）是通过改变其构象将特定的外部刺激传递到细胞中的膜蛋白。这种构象变化允许它们偶联和激活G蛋白以启动信号转导。研究和推断这些结构动力学的一个关键挑战来自细胞环境的复杂性，包括各种内源性因素的存在。由于细胞表达系统的最新进展，膜蛋白纯化技术，和标签方法，现在可以在体外和活细胞中在单分子水平上研究GPCRs的结构动力学。在这次审查中,我们讨论最先进的表达技术和策略，净化，并在单分子研究的背景下标记GPCRs。我们还重点介绍了四项最近的研究，这些研究证明了单分子显微镜在揭示GPCRs动力学中的应用。这些技术也可用作补充方法，以验证从其他结构生物学工具（如低温电子显微镜和X射线晶体学）获得的结果。
