Abstract
The assays described here are pertinent to protein kinase studies in any plant. They include an immunoblot phosphorylation/activation assay and an in-gel activity assay for MAP kinases (MPKs) using the general protein kinase substrate myelin basic protein. They also include a novel in-gel peptide substrate assay for Snf1-related kinase family 2 members (SnRK2s). This kinase family-specific assay overcomes some limitations of in-gel assays and permits the identification of different types of kinase activities in total protein extracts.
*These authors contributed equally to this work.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Fiil BK, Petersen K, Petersen M, Mundy J (2009) Gene regulation by MAP kinase cascades. Curr Opin Plant Biol 12:615–621
Rodriguez M, Petersen M, Mundy J (2010) Plant MAP kinase cascades. Annu Rev Plant Biol 61:621–649
Rasmussen M, Roux M, Petersen M, Mundy J (2012) Map kinase cascades in plant innate immunity. Front Plant Sci 3(169):1–6
Zhang S, Klessig DF (1997) Salicylic acid activates a 48-kD MAP kinase in tobacco. Plant Cell 9:809–824
Martenson RE, Law MJ, Deibler GE (1983) Identification of multiple in vivo phosphorylation site in rabbit myelin basic protein. J Biol Chem 258:930–937
Weiner JJ, Peterson FC, Volkman BF, Cutler SR (2010) Structural and functional insights into core ABA signaling. Curr Opin Plant Biol 13:495–502
Fujii H, Zhu JK (2012) Osmotic stress signaling via protein kinases. Cell Mol Life Sci 69:3165–3173
Belin C, de Franco PO, Bourbousse C, Chaignepain S, Schmitter JM, Vavasseur A, Giraudat J, Barbier-Brygoo H, Thomine S (2006) Identification of features regulating OST1 kinase activity and OST1 function in guard cells. Plant Physiol 141:1316–1327
Boudsocq M, Droillard M-J, Barbier-Brygoo H, Laurière C (2007) Different phosphorylation mechanisms are involved in the activation of sucrose non-fermenting 1 related protein kinases 2 by osmotic stresses and abscisic acid. Plant Mol Biol 63:491–503
Huang JZ, Huber SC (2001) Phosphorylation of synthetic peptides by a CDPK and plant SNF1-related protein kinase. Influence of proline and basic amino acid residues at selected positions. Plant Cell Physiol 42:1079–1087
Sirichandra C, Davanture M, Turk BE, Zivy M, Valot B, Leung J, Merlot S (2010) The Arabidopsis ABA-activated kinase OST1 phosphorylates the bZIP transcription factor ABF3 and creates a 14-3-3 binding site involved in its turnover. PLoS One 5:e13935
Dale S, Wilson WA, Edelman AM, Hardie DG (1995) Similar substrate recognition motifs for mammalian AMP-activated protein kinase, higher plant HMG-CoA reductase kinase-A, yeast SNF1, and mammalian calmodulin-dependent protein kinase I. FEBS Lett 361:191–195
Halfter U, Ishitani M, Zhu JK (2000) The Arabidopsis SOS2 protein kinase physically interacts with and is activated by the calcium-binding protein SOS3. Proc Natl Acad Sci USA 97:3735–3740
O’Brien-Simpson NM, Ede NJ, Brown LE, Swan J, Jackson DC (1997) Polymerization of unprotected synthetic peptides: a view toward synthetic peptide vaccines. J Am Chem Soc 119:1183–1188
Bressendorff S, Azevedo R, Kenchappa CS, Ponce de León I, Olsen JV, Rasmussen MW, Erbs G, Newman MA, Petersen M, Mundy J (2016) An innate immunity pathway in the moss Physcomitrella patens. Plant Cell 28:1328–1342
Yang Z, Guo G, Zhang M, Liu CY, Hu Q, Lam H, Cheng H, Xue Y, Li J, Li N (2013) Stable isotope metabolic labeling-based quantitative phosphoproteomic analysis of Arabidopsis mutants reveals ethylene-regulated time-dependent phosphoproteins and putative substrates of constitutive triple response 1 kinase. Mol Cell Proteomics 12:3559–3582
Roitinger E, Hofer M, Köcher T, Pichler P, Novatchkova M, Yang J, Schlögelhofer P, Mechtler K (2015) Quantitative phosphoproteomics of the ataxia telangiectasia-mutated (ATM) and ataxia telangiectasia-mutated and rad3-related (ATR) dependent DNA damage response in Arabidopsis thaliana. Mol Cell Proteomics 14:556–571
Jiménez C, Berl T, Rivard CJ, Edelstein CL, Capasso JM (2004) Phosphorylation of MAP kinase-like proteins mediate the response of the halotolerant alga Dunaliella viridis to hypertonic shock. Biochim Biophys Acta 1644:61–69
Romeis T, Piedras P, Zhang S, Klessig DF, Hirt H, Jones DG (1999) Rapid Avr 9- and Cf-9–dependent activation of MAP kinases in tobacco cell cultures and leaves: convergence of resistance gene, elicitor, wound, and salicylate responses. Plant Cell 11:273–288
Acknowledgments
This work was supported by grants from the Danish Basic Research Foundation (Center for Comparative Genomics) and the Danish Research Council for Nature and the Universe (Comparative & functional genomics of plant innate immunity, 1323-00267A) to JM.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Kenchappa, C. et al. (2017). Chitin and Stress Induced Protein Kinase Activation. In: Shan, L., He, P. (eds) Plant Pattern Recognition Receptors. Methods in Molecular Biology, vol 1578. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6859-6_15
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6859-6_15
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6858-9
Online ISBN: 978-1-4939-6859-6
eBook Packages: Springer Protocols