Analysis of protein-protein interactions in mitochondria by coimmunoprecipitation and chemical cross-linking

Methods Cell Biol. 2001;65:217-30. doi: 10.1016/s0091-679x(01)65013-1.

Abstract

Many different techniques have been employed to analyze protein-protein interactions. Coimmunoprecipitation and chemical cross-linking have been used extensively to study mitochondrial biogenesis. Both techniques have proven to be powerful methods to investigate the sequential interactions of precursor proteins with the various components of the translocation machineries in the mitochondrial membranes. Similarly, protein-protein interactions during processes such as protein synthesis, folding, and degradation can be studied. Moreover, the composition of the oligomeric protein complexes of mitochondria, such as respiratory chain complexes or protein translocation machineries, can be determined. The general principles and protocols of these methods are described and illustrated with typical examples.

MeSH terms

  • Adenosine Triphosphatases / chemistry
  • Adenosine Triphosphatases / metabolism
  • Cross-Linking Reagents*
  • Electron Transport Complex IV / chemistry
  • Electron Transport Complex IV / metabolism
  • Mitochondria / chemistry
  • Mitochondria / metabolism*
  • Precipitin Tests / methods*
  • Protein Subunits
  • Proteins / metabolism*
  • Tetrahydrofolate Dehydrogenase / chemistry
  • Tetrahydrofolate Dehydrogenase / metabolism

Substances

  • Cross-Linking Reagents
  • Protein Subunits
  • Proteins
  • Tetrahydrofolate Dehydrogenase
  • Electron Transport Complex IV
  • Adenosine Triphosphatases