Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, USA.
FEBS J. 2021 Oct;288(20):5788-5804. doi: 10.1111/febs.15676. Epub 2020 Dec 31.
Membrane proteins play critical physiological roles in all organisms, from ion transport and signal transduction to multidrug resistance. Elucidating their 3D structures is essential for understanding their functions, and this information can also be exploited for structure-aided drug discovery efforts. In this regard, X-ray crystallography has been the most widely used technique for determining the high-resolution 3D structures of membrane proteins. However, the success of this technique is dependent on efficient protein extraction, solubilization, stabilization, and generating diffracting crystals. Each of these steps can impose great challenges for membrane protein crystallographers. In this review, the process of generating membrane protein crystals from protein extraction and solubilization to structure determination is discussed. In addition, the current methods for precrystallization screening and a few strategies to increase the chance of crystallizing challenging membrane proteins are introduced.
膜蛋白在所有生物体中都发挥着至关重要的生理作用，从离子转运和信号转导到多药耐药性。阐明它们的三维结构对于理解其功能至关重要，这些信息也可用于基于结构的药物发现工作。在这方面，X 射线晶体学一直是确定膜蛋白高分辨率三维结构的最广泛使用的技术。然而，该技术的成功取决于高效的蛋白质提取、溶解、稳定和产生衍射晶体。这些步骤中的每一步都可能给膜蛋白晶体学家带来巨大的挑战。在这篇综述中，讨论了从蛋白质提取和溶解到结构确定的过程中生成膜蛋白晶体的过程。此外，还介绍了当前的预结晶筛选方法和一些增加结晶挑战性膜蛋白机会的策略。
