Faculty of Biology, Shenzhen MSU-BIT University, Shenzhen, China.
Faculty of Biology, Moscow Lomonosov University, Moscow, Russia.
Methods Mol Biol. 2024;2796:73-86. doi: 10.1007/978-1-0716-3818-7_4.
Structural studies require the production of target proteins in large quantities and with a high degree of purity. For membrane proteins, the bottleneck in determining their structure is the extraction of the target protein from the cell membranes. A detergent that improperly mimics the hydrophobic environment of the protein of interest can also significantly alter its structure. Recently, using lipodiscs with styrene-maleic acid (SMA), copolymers became a promising strategy for the purification of membrane proteins. Here, we describe in detail the one-step affinity purification of potassium ion channels solubilized in SMA and sample preparation for future structural studies.
结构研究需要大量且高纯度的目标蛋白。对于膜蛋白,确定其结构的瓶颈是从细胞膜中提取目标蛋白。一种去污剂如果不能正确模拟目标蛋白的疏水环境,也会显著改变其结构。最近,使用苯乙烯-马来酸(SMA)共聚物的脂筏成为一种有前途的膜蛋白纯化策略。在这里,我们详细描述了在 SMA 中溶解的钾离子通道的一步亲和纯化和未来结构研究的样品制备。