Hands on Native Mass Spectrometry Analysis of Multi-protein Complexes

Methods Mol Biol. 2021:2247:173-191. doi: 10.1007/978-1-0716-1126-5_10.

Abstract

By maintaining intact multi-protein complexes in the gas-phase, native mass spectrometry provides their molecular weight with very good accuracy compared to other methods (typically native PAGE or SEC-MALS) (Marcoux and Robinson, Structure 21:1541-1550, 2013). Besides, heterogeneous samples, in terms of both oligomeric states and ligand-bound species can be fully characterized. Here we thoroughly describe the analysis of several oligomeric protein complexes ranging from a 16 = kDa dimer to a 801-kDa tetradecameric complex on different instrumental setups.

Keywords: Noncovalent mass spectrometry; Oligomeric states; Stoichiometry; Structural mass spectrometry; Subcomplexes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, Gel
  • DNA / chemistry
  • DNA-Directed RNA Polymerases / analysis
  • DNA-Directed RNA Polymerases / chemistry
  • Escherichia coli / enzymology
  • Mass Spectrometry* / methods
  • Molecular Weight
  • Multiprotein Complexes / analysis*
  • Multiprotein Complexes / chemistry*
  • Protein Binding
  • Protein Multimerization
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Ultracentrifugation
  • Yeasts / enzymology

Substances

  • Multiprotein Complexes
  • DNA
  • DNA-Directed RNA Polymerases