Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700-8530, Japan.
STAR Protoc. 2021 Dec 16;3(1):101046. doi: 10.1016/j.xpro.2021.101046. eCollection 2022 Mar 18.
Microbial rhodopsins are photoreceptive membrane proteins showing various light-dependent biological activities. Styrene-maleic acid (SMA) copolymers spontaneously form nanoscale lipid particles containing membrane proteins and associated lipids without detergent, and can be used to characterize membrane molecules. Here, we provide a protocol to functionally express a thermally stable rhodopsin, rhodopsin, and an unstable rhodopsin, sensory rhodopsin I, in We then describe the preparation of SMA and the extraction and purification of rhodopsin molecules using SMA. For complete details on the use and execution of this protocol, please refer to Ueta et al. (2020).
微生物视紫红质是感光膜蛋白,具有各种依赖于光的生物活性。苯乙烯-马来酸(SMA)共聚物在没有去污剂的情况下自发形成含有膜蛋白和相关脂质的纳米级脂质颗粒,可用于表征膜分子。在这里,我们提供了一个功能表达热稳定视紫红质蛋白 rhodopsin 和不稳定视紫红质蛋白 sensory rhodopsin I 的方案,然后描述了 SMA 的制备以及使用 SMA 提取和纯化视紫红质分子的过程。有关该方案使用和执行的完整详细信息,请参阅 Ueta 等人(2020 年)。