title: Detecting protein-protein interactions/complex components using mass spectrometry coupled techniques.
abstract: Proteins play important roles in biochemical processes. Most biological functions are realized through protein-protein interactions (PPI). Co-immunoprecipitation is the most straightforward method to detect PPI. With the development of modern mass spectrometry (MS), throughput, sensitivity, and confidence for the detection of PPI can be readily achieved by scaling up traditional antibody-based strategies. Herein, we describe a typical workflow for general PPI detection using mass spectrometry coupled techniques, covering from Co-immunoprecipitation (Co-IP), to gel display, in-gel digestion, liquid chromatography mass spectrometry (LC-MS) analysis, as well as result interpretation and statistic filtering. This protocol provides an overview of the technique as well as practical tips.
authors.affiliation: Department of Biochemistry, Microbiology and Immunology, Ottawa Institute of Systems Biology, University of Ottawa, Roger Guindon Hall, 451 Smyth Road, Ottawa, ON, Canada, K1H 8M5, ningzhibin@gmail.com.
authors.forename: Zhibin
authors.initials: Z
authors.lastname: Ning
authors.forename: Brett
authors.initials: B
authors.lastname: Hawley
authors.forename: Cheng-Kang
authors.initials: CK
authors.lastname: Chiang
authors.forename: Deeptee
authors.initials: D
authors.lastname: Seebun
authors.forename: Daniel
authors.initials: D
authors.lastname: Figeys
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods*; D060066:Protein Interaction Maps; D011506:Proteins / Q000032:analysis / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D012984:Software
chemical_list: D011506:Proteins