Shinohara Hidefumi, Matsubayashi Yoshikatsu
Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa, Nagoya, 464-8602, Japan.
Methods Mol Biol. 2017;1621:29-35. doi: 10.1007/978-1-4939-7063-6_3.
Although more than 600 single-transmembrane receptor kinase genes have been found in the Arabidopsis genome, only a few of them have known physiological functions, and even fewer plant receptor kinases have known specific ligands. Ligand-binding analysis must be operated using the functionally expressed receptor form. However, the relative abundance of native receptor kinase molecules in the plasma membrane is often quite low. Here, we present a method for stable and functional expression of plant receptor kinases in tobacco BY-2 cells that allows preparation of microsomal fractions containing the receptor. This procedure provides a sufficient amount of receptor proteins while maintaining its ligand-binding activities.
虽然在拟南芥基因组中已发现600多个单跨膜受体激酶基因，但其中只有少数具有已知的生理功能，而植物受体激酶中已知特定配体的更是少之又少。配体结合分析必须使用功能表达的受体形式进行操作。然而，原生质膜中天然受体激酶分子的相对丰度通常相当低。在此，我们提出一种在烟草BY-2细胞中稳定且功能性表达植物受体激酶的方法，该方法可制备含有该受体的微粒体组分。此程序在保持受体配体结合活性的同时，可提供足够数量的受体蛋白。
