Pizzio Gastón A, Rodriguez Pedro L
Department of Molecular Genetics, Centre for Research in Agricultural Genomics (CRAG; consortium CSIC-IRTA-UAB-UB), Barcelona, Spain.
Instituto de Biologia Molecular y Celular de Plantas, Consejo Superior de Investigaciones Cientificas-Universidad Politecnica de Valencia, Valencia, Spain.
Bio Protoc. 2017 Mar 5;7(5):e2170. doi: 10.21769/BioProtoc.2170.
Proper spatiotemporal regulation of protein phosphorylation in cells and tissues is required for normal development and homeostasis. We present the protocol 'In Gel Kinase Assay', which is useful for protein kinase activity measurements from crude protein extracts. We have successfully used 'In Gel Kinase Assay' protocol to show that the sextuple mutant in the PYRABACTIN RESISTANCE1/PYR1-LIKE/REGULATORY COMPONENTS OF ABA RECEPTORS (PYR/PYL/RCAR-ABA receptors; line ) is impaired in ABA-mediated activation of SnRK2.2, SnRK2.3 and OST1/SnRK2.6, as much as the triple mutant (Gonzalez-Guzman , 2012).
细胞和组织中蛋白质磷酸化的适当时空调节对于正常发育和体内平衡是必需的。我们介绍了“凝胶内激酶测定”方案，该方案可用于从粗蛋白提取物中测量蛋白激酶活性。我们已成功使用“凝胶内激酶测定”方案表明，PYRABACTIN抗性1/PYR1样/ABA受体的调节成分（PYR/PYL/RCAR-ABA受体；品系）中的六重突变体在ABA介导的SnRK2.2、SnRK2.3和OST1/SnRK2.6激活方面受损，与三重突变体（Gonzalez-Guzman，2012年）一样。
