Kadota Yasuhiro, Macho Alberto P, Zipfel Cyril
The Sainsbury Laboratory, Norwich Research Park, Norwich, NR4 7UH, UK.
Plant Immunity Research Group, RIKEN Center for Sustainable Resource Science, Suehiro-cho 1-7-22 Tsurumi-ku, Yokohama, 230-0045, Japan.
Methods Mol Biol. 2016;1363:133-44. doi: 10.1007/978-1-4939-3115-6_11.
Membrane proteins are difficult to study for numerous reasons. The surface of membrane proteins is relatively hydrophobic and sometimes very unstable, additionally requiring detergents for their extraction from the membrane. This leads to challenges at all levels, including expression, solubilization, purification, identification of associated proteins, and the identification of post-translational modifications. However, recent advances in immunoprecipitation technology allow to isolate membrane proteins efficiently, facilitating the study of protein-protein interactions, the identification of novel associated proteins, and to identify post-translational modifications, such as phosphorylation. Here, we describe an optimized immunoprecipitation protocol for plant plasma membrane receptor-like kinases.
由于众多原因，膜蛋白难以研究。膜蛋白的表面相对疏水，有时非常不稳定，此外还需要去污剂才能将其从膜中提取出来。这在各个层面都带来了挑战，包括表达、溶解、纯化、相关蛋白的鉴定以及翻译后修饰的鉴定。然而，免疫沉淀技术的最新进展使得能够高效分离膜蛋白，有助于研究蛋白质 - 蛋白质相互作用、鉴定新的相关蛋白以及鉴定翻译后修饰，如磷酸化。在此，我们描述了一种针对植物质膜类受体激酶的优化免疫沉淀方案。
