title: Immunoprecipitation of Plasma Membrane Receptor-Like Kinases for Identification of Phosphorylation Sites and Associated Proteins.
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.
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich, NR4 7UH, UK.|Plant Immunity Research Group, RIKEN Center for Sustainable Resource Science, Suehiro-cho 1-7-22 Tsurumi-ku, Yokohama, 230-0045, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kadota
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich, NR4 7UH, UK.|Shanghai Center for Plant Stress Biology, Chinese Academy of Sciences, Shanghai, 201602, China.
authors.forename: Alberto P
authors.initials: AP
authors.lastname: Macho
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich, NR4 7UH, UK. cyril.zipfel@tsl.ac.uk.
authors.forename: Cyril
authors.initials: C
authors.lastname: Zipfel
mesh_terms: D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D047468:Immunoprecipitation / Q000379:methods; D008565:Membrane Proteins / Q000378:metabolism*; D010766:Phosphorylation; D010940:Plant Proteins / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D008565:Membrane Proteins; D010940:Plant Proteins; D011494:Protein Kinases
keywords: Immunoprecipitation; Protein phosphorylation; Receptor-like kinase