Schweighofer Alois, Shubchynskyy Volodymyr, Kazanaviciute Vaiva, Djamei Armin, Meskiene Irute
Max F. Perutz Laboratories, University and Medical University of Vienna, Dr. Bohrgasse 9, 1030, Vienna, Austria,
Methods Mol Biol. 2014;1171:147-58. doi: 10.1007/978-1-4939-0922-3_12.
The adaptation of plants to the environment is a key property for survival. Adaptation responses to environmental cues are generated in cells by signaling initiated from cell receptors. Signal transduction is based on protein phosphorylation that is employed in mitogen-activated protein kinase (MAPK) cascades to integrate signals from receptors to cellular responses. MAPK activity is determined by phosphorylation of amino acid residues within the kinase activation loop and their dephosphorylation by phosphatases is essential to control signal duration and intensity.Monitoring protein-protein interactions (PPIs) of MAPKs with MAPK phosphatases in vivo provides valuable information about specificity and intracellular localization of the protein complex. Here, we report studying PPIs between Arabidopsis MAPKs and PP2C-type MAPK phosphatases using bimolecular fluorescent complementation (BiFC) in suspension cell protoplasts. The interactions of the MAPKs MPK3, MKP4 and MPK6 with the phosphatases AP2C1 and AP2C3 have been tested.
植物对环境的适应性是其生存的关键特性。细胞通过细胞受体启动的信号传导产生对环境信号的适应性反应。信号转导基于蛋白质磷酸化，其在丝裂原活化蛋白激酶（MAPK）级联反应中用于将受体信号整合到细胞反应中。MAPK活性由激酶激活环内氨基酸残基的磷酸化决定，而磷酸酶对其去磷酸化对于控制信号持续时间和强度至关重要。在体内监测MAPK与MAPK磷酸酶之间的蛋白质-蛋白质相互作用（PPI）可提供有关蛋白质复合物特异性和细胞内定位的有价值信息。在此，我们报告了在悬浮细胞原生质体中使用双分子荧光互补（BiFC）研究拟南芥MAPK与PP2C型MAPK磷酸酶之间的PPI。已经测试了MAPK MPK3、MKP4和MPK6与磷酸酶AP2C1和AP2C3之间的相互作用。
