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Mass Photometry of Membrane Proteins

Olerinyova, Anna and Sonn-Segev, Adar and Gault, Joseph and Eichmann, Cédric and Schimpf, Johannes and Kopf, Adrian H. and Rudden, Lucas S.P. and Ashkinadze, Dzmitry and Bomba, Radoslaw and Frey, Lukas and Greenwald, Jason and Degiacomi, Matteo T. and Steinhilper, Ralf and Killian, J. Antoinette and Friedrich, Thorsten and Riek, Roland and Struwe, Weston B. and Kukura, Philipp (2021) 'Mass Photometry of Membrane Proteins.', Chem., 7 (1). pp. 224-236.

Abstract

Integral membrane proteins (IMPs) are biologically highly significant but challenging to study because they require maintaining a cellular lipid-like environment. Here, we explore the application of mass photometry (MP) to IMPs and membrane-mimetic systems at the single-particle level. We apply MP to amphipathic vehicles, such as detergents and amphipols, as well as to lipid and native nanodiscs, characterizing the particle size, sample purity, and heterogeneity. Using methods established for cryogenic electron microscopy, we eliminate detergent background, enabling high-resolution studies of membrane-protein structure and interactions. We find evidence that, when extracted from native membranes using native styrene-maleic acid nanodiscs, the potassium channel KcsA is present as a dimer of tetramers—in contrast to results obtained using detergent purification. Finally, using lipid nanodiscs, we show that MP can help distinguish between functional and non-functional nanodisc assemblies, as well as determine the critical factors for lipid nanodisc formation.

Item Type:Article
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Available under License - Creative Commons Attribution Non-commercial No Derivatives 4.0.
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Status:Peer-reviewed
Publisher Web site:https://doi.org/10.1016/j.chempr.2020.11.011
Publisher statement:© 2020 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Date accepted:11 November 2020
Date deposited:04 May 2021
Date of first online publication:14 December 2020
Date first made open access:04 May 2021

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