Structure Determination of Membrane Proteins Using X-Ray Crystallography.
Structure Determination of Membrane Proteins Using X-Ray Crystallography.
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DOI：
10.1007/978-1-0716-1394-8_7
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Noinaj N
中科院分区：
文献类型：
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作者：
Billings E;Lundquist K;Overly C;Srinivasan K;Noinaj N
膜蛋白在生命的各个方面发挥着重要作用，约占从原核生物到真核生物所有基因组的三分之一。它们的职责包括介导细胞信号传导、营养输入、废物输出、细胞通信、贩运和免疫。由于它们在许多细胞过程中的关键作用，膜蛋白作为目前市场上高达50%的药物的靶标，并且仍然是正在开发的新疗法的主要靶标。尽管它们在自然界中的重要性和丰富性，但在蛋白质数据库中只有约1%的结构是跨膜蛋白。这种差异可以直接归因于膜蛋白的生物化学性质以及难以产生足够的产量用于结构研究或难以生长有序晶体。在这里，我们介绍了我们工作中的方法，概述了我们从克隆到膜蛋白结构测定的一般管道，重点是使用X射线晶体学，它仍然产生约90%的结构被存入蛋白质数据库。
Membrane proteins serve essential roles in all aspects of life and make up roughly one-third of all genomes from prokaryotes to eukaryotes. Their responsibilities include mediating cell signaling, nutrient import, waste export, cellular communication, trafficking, and immunity. For their critical role in many cellular processes, membrane proteins serve as targets for up to 50% of drugs currently on the market and remain primary targets in new therapeutics being developed. Despite their importance and abundance in nature, only ~1% of structures in the Protein Data Bank are of transmembrane proteins. This discrepancy can be directly attributed to the biochemical properties of membrane proteins and the difficulty in producing sufficient yields for structural studies or the difficulty in growing well-ordered crystals. Here, we present methods from our work that outline our general pipeline from cloning to structure determination of membrane proteins, with a focus on using X-ray crystallography, which still yields ~90% of all structures being deposited into the Protein Data Bank.