Global Characterization of Protein Complexes by Biochemical Purification-Mass Spectrometry (BP/MS)

Reza Pourhaghighi; Andrew Emili

doi:10.1007/978-1-4939-7759-8_12

Global Characterization of Protein Complexes by Biochemical Purification-Mass Spectrometry (BP/MS)

Methods Mol Biol. 2018:1764:185-191. doi: 10.1007/978-1-4939-7759-8_12.

Authors

Reza Pourhaghighi¹, Andrew Emili^{2

3}

Affiliations

¹ Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON, Canada.
² Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON, Canada. andrew.emili@utoronto.ca.
³ Departments of Biology and Biochemistry, Boston University, Toronto, ON, Canada. andrew.emili@utoronto.ca.

PMID: 29605916
DOI: 10.1007/978-1-4939-7759-8_12

Abstract

A proteomic platform for global analysis of protein complexes and protein-protein interactions (PPIs) is described. Briefly, after comprehensive physiochemical separation of soluble protein extracts using non-denaturing ion exchange chromatography (IEX), each fraction is subjected to quantitative tandem mass spectrometry analysis.

Keywords: High Performance Liquid Chromatography (HPLC); Intact Protein Separation; Ion Exchange Chromatography (IEX); Nano-Liquid Chromatography Tandem Mass Spectrometry (nLC-MS/MS); Protein-Protein Interaction.

MeSH terms

Chromatography, High Pressure Liquid
Chromatography, Ion Exchange / methods*
Humans
Protein Binding
Protein Interaction Domains and Motifs*
Proteins / chemistry
Proteins / isolation & purification*
Proteins / metabolism*
Proteomics / methods*
Tandem Mass Spectrometry / methods*

Substances

Proteins