Mass spectrometry of intact membrane protein complexes

Arthur Laganowsky, Eamonn Reading, Jonathan T S Hopper, Carol V Robinson

Research output: Contribution to journalArticlepeer-review

280 Citations (Scopus)
62 Downloads (Pure)


Mass spectrometry (MS) of intact soluble protein complexes has emerged as a powerful technique to study the stoichiometry, structure-function and dynamics of protein assemblies. Recent developments have extended this technique to the study of membrane protein complexes, where it has already revealed subunit stoichiometries and specific phospholipid interactions. Here we describe a protocol for MS of membrane protein complexes. The protocol begins with the preparation of the membrane protein complex, enabling not only the direct assessment of stoichiometry, delipidation and quality of the target complex but also the evaluation of the purification strategy. A detailed list of compatible nonionic detergents is included, along with a protocol for screening detergents to find an optimal one for MS, biochemical and structural studies. This protocol also covers the preparation of lipids for protein-lipid binding studies and includes detailed settings for a quadrupole time-of-flight (Q-TOF) mass spectrometer after the introduction of complexes from gold-coated nanoflow capillaries.

Original languageEnglish
Pages (from-to)639-51
Number of pages13
JournalNature Protocols
Issue number4
Publication statusPublished - Apr 2013


  • Detergents/chemistry
  • Indicators and Reagents/chemistry
  • Mass Spectrometry/instrumentation
  • Membrane Proteins/chemistry
  • Phospholipids/chemistry
  • Protein Binding
  • Protein Structure, Tertiary


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