Charge Reduction of Membrane Proteins in Native Mass Spectrometry Using Alkali Metal Acetate Salts

Anal Chem. 2020 May 5;92(9):6622-6630. doi: 10.1021/acs.analchem.0c00454. Epub 2020 Apr 14.

Abstract

Native mass spectrometry (MS) provides the capacity to monitor membrane protein complexes and noncovalent binding of ligands and lipids to membrane proteins. The charge states produced by native MS of membrane proteins often result in gas-phase protein unfolding or loss of noncovalent interactions. In an effort to reduce the charge of membrane proteins, we examined the utility of alkali metal salts as a charge-reducing agent. Low concentrations of alkali metal salts caused marked charge reduction in the membrane protein, Erwinia ligand-gated ion channel (ELIC). The charge-reducing effect only occurred for membrane proteins and was detergent-dependent, being most pronounced in long polyethylene glycol (PEG)-based detergents such as C10E5 and C12E8. On the basis of these results, we propose a mechanism for alkali metal charge reduction of membrane proteins. Addition of low concentrations of alkali metals may provide an advantageous approach for charge reduction of detergent-solubilized membrane proteins by native MS.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Acetates / chemistry*
  • Animals
  • Cattle
  • Detergents / chemistry
  • Glutamate Dehydrogenase / chemistry*
  • Glutamate Dehydrogenase / metabolism
  • Mass Spectrometry
  • Membrane Proteins / chemistry*
  • Membrane Proteins / metabolism
  • Metals, Alkali / chemistry*
  • Oxidation-Reduction
  • Pyruvate Kinase / chemistry*
  • Pyruvate Kinase / metabolism
  • Rabbits
  • Salts / chemistry
  • Solubility

Substances

  • Acetates
  • Detergents
  • Membrane Proteins
  • Metals, Alkali
  • Salts
  • Glutamate Dehydrogenase
  • Pyruvate Kinase