Immunoprecipitation of Plasma Membrane Receptor-Like Kinases for Identification of Phosphorylation Sites and Associated Proteins

  • Yasuhiro Kadota
  • Alberto P. Macho
  • Cyril ZipfelEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1363)


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.

Key words

Immunoprecipitation Protein phosphorylation Receptor-like kinase 



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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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Yasuhiro Kadota
    • 1
    • 2
  • Alberto P. Macho
    • 1
    • 3
  • Cyril Zipfel
    • 1
    Email author
  1. 1.The Sainsbury LaboratoryNorwich Research ParkNorwichUK
  2. 2.Plant Immunity Research GroupRIKEN Center for Sustainable Resource ScienceYokohamaJapan
  3. 3.Shanghai Center for Plant Stress BiologyChinese Academy of SciencesShanghaiChina

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