Watkinson, TG, Calabrese, AN, Giusti, F et al. (3 more authors) (2015) Systematic analysis of the use of amphipathic polymers for studies of outer membrane proteins using mass spectrometry. International Journal of Mass Spectrometry, 391. pp. 54-61. ISSN 1387-3806
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.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2015, The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
Keywords: | Outer membrane proteins; amphipols; protein conformation; electrospray ionisation–mass spectrometry; ion mobility spectrometry–mass spectrometry |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 05 Nov 2015 09:58 |
Last Modified: | 12 Feb 2019 13:46 |
Published Version: | http://dx.doi.org/10.1016/j.ijms.2015.06.017 |
Status: | Published |
Publisher: | Elsevier |
Identification Number: | 10.1016/j.ijms.2015.06.017 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:91435 |