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Mass Photometry of Membrane Proteins

Olerinyova, Anna; Sonn-Segev, Adar; Gault, Joseph; Eichmann, Cédric; Schimpf, Johannes; Kopf, Adrian H.; Rudden, Lucas S.P.; Ashkinadze, Dzmitry; Bomba, Radoslaw; Frey, Lukas; Greenwald, Jason; Degiacomi, Matteo T.; Steinhilper, Ralf; Killian, J. Antoinette; Friedrich, Thorsten; Riek, Roland; Struwe, Weston B.; Kukura, Philipp

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Authors

Anna Olerinyova

Adar Sonn-Segev

Joseph Gault

Cédric Eichmann

Johannes Schimpf

Adrian H. Kopf

Lucas S.P. Rudden

Dzmitry Ashkinadze

Radoslaw Bomba

Lukas Frey

Jason Greenwald

Ralf Steinhilper

J. Antoinette Killian

Thorsten Friedrich

Roland Riek

Weston B. Struwe

Philipp Kukura



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.

Citation

Olerinyova, A., Sonn-Segev, A., Gault, J., Eichmann, C., Schimpf, J., Kopf, A. H., …Kukura, P. (2021). Mass Photometry of Membrane Proteins. Chem, 7(1), 224-236. https://doi.org/10.1016/j.chempr.2020.11.011

Journal Article Type Article
Acceptance Date Nov 11, 2020
Online Publication Date Dec 14, 2020
Publication Date Jan 14, 2021
Deposit Date Dec 21, 2020
Publicly Available Date May 4, 2021
Journal Chem
Print ISSN 2451-9308
Publisher Cell Press
Peer Reviewed Peer Reviewed
Volume 7
Issue 1
Pages 224-236
DOI https://doi.org/10.1016/j.chempr.2020.11.011
Public URL https://durham-repository.worktribe.com/output/1255110
Related Public URLs https://ora.ox.ac.uk/objects/uuid:edb7aa7b-e5e2-4c83-b279-b7a4d056ccef

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