title: CDPK Activation in PRR Signaling.
abstract: Calcium-dependent protein kinases undergo a rapid biochemical activation in response to an intracellular Ca increase induced by the PRR-dependent perception of a pathogen-related stimulus. Based on SDS gel resolution, the in-gel kinase assay allows the analysis of multiple in vivo protein samples in parallel, combining the advantage of protein separation according to molecular mass with the activity read-out of a protein kinase assay. It thus enables to follow the transient CDPK activation and inactivation in response to in vivo elicitation with a time-wise resolution. In addition, changes of CDPK phosphorylation activity often correlate with slight shifts in the enzyme's apparent molecular mass, indicating posttranslational modifications and a conformational change of the active enzyme compared to its inactive resting form. These band shifts can be detected by a simple immunoblotting to monitor CDPK activation.
authors.affiliation: Dahlem Centre of Plant Sciences, Plant Biochemistry, Freie Universität Berlin, Königin-Luise-Straße 12-16, 14195, Berlin, Germany.
authors.forename: Heike
authors.initials: H
authors.lastname: Seybold
authors.affiliation: Institute of Plant Sciences Paris-Saclay (IPS2), CNRS, INRA, Université Paris-Sud, Université d'Evry Val d'Essonne, Université Paris-Diderot, Sorbonne Paris-Cité, Université Paris-Saclay, Bâtiment 630, 91405, Orsay, France. marie.boudsocq@ips2.universite-paris-saclay.fr.
authors.forename: Marie
authors.initials: M
authors.lastname: Boudsocq
authors.affiliation: Dahlem Centre of Plant Sciences, Plant Biochemistry, Freie Universität Berlin, Königin-Luise-Straße 12-16, 14195, Berlin, Germany.
authors.forename: Tina
authors.initials: T
authors.lastname: Romeis
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D024202:Electrophoretic Mobility Shift Assay; D004789:Enzyme Activation; D008970:Molecular Weight; D010766:Phosphorylation; D011487:Protein Conformation; D011494:Protein Kinases / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; D014157:Transcription Factors; D011494:Protein Kinases
keywords: CDPK; Electrophoretic band shift; In-gel kinase assay; Phosphorylation target; Protoplast; Signal transduction; Transient expression