Mitochondrial Complexome Profiling

Methods Mol Biol. 2021:2192:269-285. doi: 10.1007/978-1-0716-0834-0_19.

Abstract

Complexome profiling combines blue native gel electrophoresis (BNE) and quantitative mass spectrometry to define an entire protein interactome of a cell, an organelle, or a biological membrane preparation. The method allows the identification of protein assemblies with low abundance and detects dynamic processes of protein complex assembly. Applications of complexome profiling range from the determination of complex subunit compositions, assembly of single protein complexes, and supercomplexes to comprehensive differential studies between patients or disease models. This chapter describes the workflow of complexome profiling from sample preparation, mass spectrometry to data analysis with a bioinformatics tool.

Keywords: Assembly; Blue native electrophoresis; Complexome profiling; Mass spectrometry; Membrane protein complexes; Mitochondria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Chromatography, Liquid / methods
  • Humans
  • Mass Spectrometry / methods*
  • Membrane Proteins / chemistry
  • Membrane Proteins / metabolism
  • Mitochondria / chemistry*
  • Mitochondria / metabolism
  • Mitochondrial Membranes / chemistry
  • Mitochondrial Membranes / metabolism
  • Mitochondrial Proteins / chemistry
  • Mitochondrial Proteins / metabolism
  • Native Polyacrylamide Gel Electrophoresis / methods*
  • Peptides / chemistry
  • Peptides / metabolism
  • Tandem Mass Spectrometry / methods

Substances

  • Membrane Proteins
  • Mitochondrial Proteins
  • Peptides