Abstract
Identification of a particular protein as a physiological substrate towards kinases of interest is an extremely complex process. Under physiological conditions kinases and their putative substrates are in low abundance, and production of active eukaryotic kinases with standard overexpressing methods is an arduous task. Herein, we describe a cell-free in vitro protein translation procedure combined with fluorescent phosphoprotein staining as a simple and rapid method for identification of putative kinase substrates.
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Acknowledgments
This work was supported by the Hungarian Scientific Research Fund and National Office for Research and Technology grant K69187 and K68160. T.M. gratefully acknowledges the support of Bolyai János fellowship.
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Nagy, S.K., Mészáros, T. (2014). In Vitro Translation-Based Protein Kinase Substrate Identification. In: Alexandrov, K., Johnston, W. (eds) Cell-Free Protein Synthesis. Methods in Molecular Biology, vol 1118. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-782-2_15
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DOI: https://doi.org/10.1007/978-1-62703-782-2_15
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-781-5
Online ISBN: 978-1-62703-782-2
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