Abstract
The highly conserved nature of the protein kinase catalytic domain and the low permeability of plant cell membranes pose a challenge to the development of specific inhibitors that target individual protein kinases in vivo. Here, we describe a chemical-genetic approach to specifically sensitize individual plant kinases to cell-permeable small molecules that do not inhibit wild-type kinases. In this approach, a single amino-acid substitution is introduced in the ATP-binding site of the enzyme enabling specific binding of ATP-competitive molecules. Cell-permeable molecules can then be used to specifically target the sensitized allele in transgenic Arabidopsis thaliana plants that do not express the wild-type form of the kinase. This strategy provides a useful tool for the functional characterization of protein kinases in planta and for the dissection of the signaling pathways in which they are involved.
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Acknowledgments
We thank Tal Sherman for his advice on Arabidopsis plant transformations. This work was supported by the US–Israel Binational Agricultural Research and Development Fund (BARD; grant no. IS-4159-08C to G.S.) and by the Binational Science Foundation (BSF; grant no. 2007091 to G.S. and K.M.S.).
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Salomon, D., Zhang, C., Shokat, K.M., Sessa, G. (2011). Sensitizing Plant Protein Kinases to Specific Inhibition by ATP-Competitive Molecules. In: Dissmeyer, N., Schnittger, A. (eds) Plant Kinases. Methods in Molecular Biology, vol 779. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-61779-264-9_10
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DOI: https://doi.org/10.1007/978-1-61779-264-9_10
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