Membrane Structural and Functional Biology Group, School of Medicine and School of Biochemistry and Immunology, Trinity College Dublin, Dublin, Ireland.
Methods Mol Biol. 2022;2507:241-271. doi: 10.1007/978-1-0716-2368-8_13.
G protein-coupled receptors (GPCRs) play vital roles in human physiology and pathophysiology. This makes the elucidation of the high-resolution blueprints of these high value membrane proteins of crucial importance for the structure-based design of novel therapeutics. However, the production and crystallization of GPCRs for structure determination comes with many challenges.In this chapter, we provide a comprehensive protocol for expressing and purifying the thromboxane A receptor (TPR), an attractive therapeutic target, for use in structure studies. Guidelines for crystallizing the TPR are also included. Together, these procedures provide a template for generating crystal structures of the TPR and indeed other GPCRs in complex with pharmacologically interesting ligands.
G 蛋白偶联受体（GPCRs）在人类生理学和病理生理学中发挥着重要作用。这使得阐明这些高价值膜蛋白的高分辨率蓝图对于基于结构的新型治疗药物设计至关重要。然而，为了进行结构测定而生产和结晶 GPCRs 存在许多挑战。在本章中，我们提供了一种全面的方案，用于表达和纯化血栓烷 A 受体（TPR），这是一个有吸引力的治疗靶点，可用于结构研究。还包括了结晶 TPR 的指南。这些程序一起为生成 TPR 甚至其他与药理学上有趣的配体结合的 GPCR 的晶体结构提供了模板。
