Identification of Associated Proteins by Immunoprecipitation and Mass Spectrometry Analysis.
Protein-protein interactions play central roles in intercellular and intracellular signal transduction. Impairment of protein-protein interactions causes many diseases such as cancer, cardiomyopathies, diabetes, microbial infections, and genetic and neurodegenerative disorders. Immunoprecipitation is a technique in which a target protein of interest bound by an antibody is used to pull down the protein complex out of cell lysates, which can be identified by mass spectrometry. Here, we describe the protocol to immunoprecipitate and identify the components of the protein complexes of ElmoE in Dictyostelium discoideum cells.
Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, South Chongqing Road No. 280, Shanghai, 200025, China.
Xiumei
X
Cao
Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, South Chongqing Road No. 280, Shanghai, 200025, China. yanjianshe@sjtu.edu.cn.
Jianshe
J
Yan
D002478:Cells, Cultured; D002853:Chromatography, Liquid; D000242:Cyclic AMP / Q000378:metabolism; D004023:Dictyostelium / Q000378:metabolism; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D025941:Protein Interaction Mapping / Q000379:methods; D053719:Tandem Mass Spectrometry
D000242:Cyclic AMP
Dictyostelium discoideum; Immunoprecipitation; Mass spectrometry; Protein-protein interaction