Systematic analysis of the use of amphipathic polymers for studies of outer membrane proteins using mass spectrometry

Thomas G. Watkinson, Antonio N. Calabrese, Fabrice Giusti, Manuela Zoonens, Sheena E. Radford, Alison E. Ashcroft

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

Membrane proteins (MPs) are essential for numerous important biological processes. Recently, mass spectrometry (MS), coupled with an array of related techniques, has been used to probe the structural properties of MPs and their complexes. Typically, detergent micelles have been employed for delivering MPs into the gas-phase, but these complexes have intrinsic properties that can limit the utility of structural studies of MPs using MS methods. Amphipols (APols) have advantages over detergent micelles and have been shown to be capable of delivering native MPs into the gas-phase. Comparing six different APols which vary in mass and charge, and the detergent n-dodecyl-β-d-maltopyranoside, we aimed to determine which APols are most efficient for delivery of native outer membrane proteins (OMPs) into the gas-phase. We show that maintaining the solution-phase folding and global structures of three different OMPs (PagP, OmpT and tOmpA) are independent of the APol used, but differences in OMP activity can result from the different APol:OMP complexes. ESI-IMS-MS analysis of OMP:APol complexes shows that the A8-35 APol is most proficient at liberating all three OMPs into the gas-phase, without altering their gas-phase conformations.

Original languageEnglish
Pages (from-to)54-61
Number of pages8
JournalInternational Journal of Mass Spectrometry
Volume391
Early online date7 Jul 2015
DOIs
Publication statusPublished - 30 Nov 2015

Keywords

  • Amphipols
  • Electrospray ionisation-mass spectrometry
  • Ion mobility spectrometry-mass spectrometry
  • Outer membrane proteins
  • Protein conformation

ASJC Scopus subject areas

  • Instrumentation
  • Condensed Matter Physics
  • Spectroscopy
  • Physical and Theoretical Chemistry

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