Leney, AC, McMorran, LM, Radford, SE et al. (1 more author) (2012) Amphipathic polymers enable the study of functional membrane proteins in the gas phase. Analytical Chemistry, 84 (22). 9841 - 9847. ISSN 1086-4377
Abstract
Membrane proteins are notoriously challenging to analyze using mass spectrometry (MS) because of their insolubility in aqueous solution. Current MS methods for studying intact membrane proteins involve solubilization in detergent. However, detergents can destabilize proteins, leading to protein unfolding and aggregation, or resulting in inactive entities. Amphipathic polymers, termed amphipols, can be used as a substitute for detergents and have been shown to enhance the stability of membrane proteins. Here, we show the utility of amphipols for investigating the structural and functional properties of membrane proteins using electrospray ionization mass spectrometry (ESI-MS). The functional properties of two bacterial outer-membrane β-barrel proteins, OmpT and PagP, in complex with the amphipol A8-35 are demonstrated, and their structural integrities are confirmed in the gas phase using ESI-MS coupled with ion mobility spectrometry (IMS). The data illustrate the power of ESI-IMS-MS in separating distinct populations of amphipathic polymers from the amphipol-membrane complex while maintaining a conformationally "nativelike" membrane protein structure in the gas phase. Together, the data indicate the potential importance and utility of amphipols for the analysis of membrane proteins using MS.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Keywords: | Acyltransferases; bacterial proteins; escherichia coli proteins; gases; hydrophobic and hydrophilic interactions; models, molecular; polymers; porins; protein folding; protein structure, secondary; spectrometry, mass, electrospray ionization |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 18 Feb 2015 11:21 |
Last Modified: | 23 Oct 2015 16:20 |
Published Version: | http://dx.doi.org/10.1021/ac302223s |
Status: | Published |
Publisher: | American Chemical Society |
Identification Number: | 10.1021/ac302223s |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:83113 |