Division of Biology and Chemistry, Laboratory of Biomolecular Research, Paul Scherrer Institute, Villigen PSI, Switzerland.
Methods Mol Biol. 2020;2127:321-338. doi: 10.1007/978-1-0716-0373-4_21.
Membrane proteins are highly interesting targets due to their pivotal role in cell function and disease. They are inserted in cell membranes, are often intrinsically flexible, and can adopt several conformational states to carry out their function. Although most overall folds of membrane proteins are known, many questions remain about specific functionally relevant intramolecular rearrangements that require experimental structure determination. Here, using the example of rhodopsin, we describe how to prepare and analyze membrane protein crystals for serial crystallography at room temperature, a new technique allowing to merge diffraction data from thousands of injector-delivered crystals that are too tiny for classical single-crystal analysis even in cryogenic conditions. The application of serial crystallography for studying protein dynamics is mentioned.
膜蛋白因其在细胞功能和疾病中的关键作用而成为极具研究价值的靶点。它们被插入细胞膜中，通常具有内在的灵活性，并可以采用多种构象状态来发挥其功能。尽管大多数膜蛋白的整体折叠结构已经为人所知，但仍有许多关于特定功能相关的分子内重排的问题有待解决，这些重排需要通过实验结构测定来确定。在这里，我们以视紫红质为例，描述了如何制备和分析膜蛋白晶体，以便在室温下进行连续结晶学研究，这是一种新技术，可将来自数千个注入器输送的晶体的衍射数据合并，即使在低温条件下，对于经典的单晶分析来说，这些晶体也太小了。文中还提到了连续结晶学在研究蛋白质动力学方面的应用。
