Identification of Associated Proteins by Immunoprecipitation and Mass Spectrometry Analysis

Methods Mol Biol. 2016:1407:131-9. doi: 10.1007/978-1-4939-3480-5_10.

Abstract

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.

Keywords: Dictyostelium discoideum; Immunoprecipitation; Mass spectrometry; Protein-protein interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Chromatography, Liquid
  • Cyclic AMP / metabolism
  • Dictyostelium / metabolism
  • Immunoprecipitation*
  • Mass Spectrometry*
  • Protein Interaction Mapping* / methods
  • Tandem Mass Spectrometry

Substances

  • Cyclic AMP