Fischer Cornelia, Sauter Margret, Dietrich Petra
Molecular Plant Physiology, Department of Biology, University of Erlangen-Nuremberg, Staudtstrasse 5, 91058, Erlangen, Germany.
Plant Developmental Biology and Plant Physiology, University of Kiel, Am Botanischen Garten 5, 24118, Kiel, Germany.
Methods Mol Biol. 2017;1621:141-149. doi: 10.1007/978-1-4939-7063-6_14.
Plant receptor-like kinases (RLKs) are regulated at various levels including posttranscriptional modification and interaction with regulatory proteins. Calmodulin (CaM) is a calcium-sensing protein that was shown to bind to some RLKs such as the PHYTOSULFOKINE RECEPTOR1 (PSKR1). The CaM-binding site is embedded in subdomain VIa of the kinase domain. It is possible that many more of RLKs interact with CaM than previously described. To unequivocally confirm CaM binding, several methods exist. Bimolecular fluorescence complementation (BiFC) and pull-down assays have been successfully used to study CaM binding to PSKR1 and are described in this chapter (BiFC) and in Chapter 15 (pull down). The two methods are complementary. BiFC is useful to show localization and interaction of soluble as well as of membrane-bound proteins in planta.
植物类受体激酶（RLK）在包括转录后修饰以及与调节蛋白相互作用等多个层面受到调控。钙调蛋白（CaM）是一种钙传感蛋白，已证明它能与一些RLK结合，如植物硫肽激素受体1（PSKR1）。CaM结合位点嵌入激酶结构域的VIa亚结构域中。可能有比先前描述的更多的RLK与CaM相互作用。为了明确证实CaM结合，有几种方法。双分子荧光互补（BiFC）和下拉分析已成功用于研究CaM与PSKR1的结合，本章（BiFC）和第15章（下拉分析）对此进行了描述。这两种方法是互补的。BiFC有助于显示植物中可溶性以及膜结合蛋白的定位和相互作用。
