In Gel Kinase Assay

Gastón A Pizzio; Pedro L Rodriguez

doi:10.21769/BioProtoc.2170

In Gel Kinase Assay

Bio Protoc. 2017 Mar 5;7(5):e2170. doi: 10.21769/BioProtoc.2170.

Authors

Gastón A Pizzio¹, Pedro L Rodriguez²

Affiliations

¹ Department of Molecular Genetics, Centre for Research in Agricultural Genomics (CRAG; consortium CSIC-IRTA-UAB-UB), Barcelona, Spain.
² Instituto de Biologia Molecular y Celular de Plantas, Consejo Superior de Investigaciones Cientificas-Universidad Politecnica de Valencia, Valencia, Spain.

Abstract

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 Arabidopsis thaliana sextuple mutant in the PYRABACTIN RESISTANCE1/PYR1-LIKE/REGULATORY COMPONENTS OF ABA RECEPTORS (PYR/PYL/RCAR-ABA receptors; line pyr/pyl112458) is impaired in ABA-mediated activation of SnRK2.2, SnRK2.3 and OST1/SnRK2.6, as much as the triple mutant snrk2.2/2.3/2.6 (Gonzalez-Guzman et al., 2012).

Keywords: ABA; Arabidopsis; Enzymatic activity; Kinases; Phosphorylation; SnRKs; in vivo.