Lee Jin Suk
Department of Biology, Concordia University, 7141 Sherbrooke W., Montreal, QC, Canada, H4B 1R6.
Methods Mol Biol. 2017;1621:47-56. doi: 10.1007/978-1-4939-7063-6_5.
Receptor kinases play a central role in various biological processes, but due to their low abundance and highly hydrophobic and dynamic nature, only a few of them have been functionally characterized, and their partners and ligands remain unidentified. Receptor protein extraction and purification from plant tissues is one of the most challenging steps for the success of various biochemical analyses to characterize their function. Immunoprecipitation is a widely used and selective method for enriching or purifying a specific protein. Here we describe two different optimized protein purification protocols, batch and on-chip immunoprecipitation, which efficiently isolate plant membrane receptor kinases for functional analysis.
受体激酶在各种生物过程中发挥着核心作用，但由于它们丰度低、高度疏水且具有动态特性，只有少数受体激酶的功能得到了表征，其相互作用蛋白和配体仍未明确。从植物组织中提取和纯化受体蛋白是各种生化分析成功表征其功能最具挑战性的步骤之一。免疫沉淀是一种广泛使用的选择性富集或纯化特定蛋白质的方法。在此，我们描述了两种不同的优化蛋白质纯化方案，即批量免疫沉淀和芯片上免疫沉淀，它们能有效分离植物膜受体激酶用于功能分析。
