Abstract
Membrane proteins are difficult to study for numerous reasons. The surface of membrane proteins is relatively hydrophobic and sometimes very unstable, additionally requiring detergents for their extraction from the membrane. This leads to challenges at all levels, including expression, solubilization, purification, identification of associated proteins, and the identification of post-translational modifications. However, recent advances in immunoprecipitation technology allow to isolate membrane proteins efficiently, facilitating the study of protein-protein interactions, the identification of novel associated proteins, and to identify post-translational modifications, such as phosphorylation. Here, we describe an optimized immunoprecipitation protocol for plant plasma membrane receptor-like kinases.
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Acknowledgements
We thank Sacco de Vries for sharing initial advices on the co-immunoprecipitation protocol, and past and present members of the Zipfel laboratory for discussion and advices. The excellent work of the TSL Proteomics support group is also acknowledged. This work was supported by the Gatsby Charitable Foundation (C.Z.) and the European Research Council (C.Z.). Fellowships from RIKEN Special Postdoctoral Research Fellowship, Excellent Young Researcher Overseas Visit Program, and Uehara Memorial Foundation to Y.K. are acknowledged. A.P.M. was funded by a long-term fellowship from the Federation of European Biochemical Societies.
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Kadota, Y., Macho, A.P., Zipfel, C. (2016). Immunoprecipitation of Plasma Membrane Receptor-Like Kinases for Identification of Phosphorylation Sites and Associated Proteins. In: Botella, J., Botella, M. (eds) Plant Signal Transduction. Methods in Molecular Biology, vol 1363. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3115-6_11
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DOI: https://doi.org/10.1007/978-1-4939-3115-6_11
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3114-9
Online ISBN: 978-1-4939-3115-6
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