Mass photometry of membrane proteins

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.
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 styrenemaleic
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

Angaben zum Objekt

• Standort: Deutsche Nationalbibliothek Frankfurt am Main
• Umfang: Online-Ressource
• Sprache: Englisch
• Anmerkungen: Chem. - 7, 1 (2021) , 224-236, ISSN: 2451-9294

Beteiligte, Orts- und Zeitangaben

• Ereignis: Veröffentlichung
• (wo): Freiburg
• (wer): Universität
• (wann): 2022
• Urheber: 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 Thomas ; Steinhilper, Ralf ; Friedrich, Thorsten ; Riek, Roland ; Struwe, Weston B. ; Kukura, Philipp
• Beteiligte Personen und Organisationen: Institute of Biochemistry, Molecular Bioenergetics

Weitere Informationen

• DOI: 10.1016/j.chempr.2020.11.011
• URN: urn:nbn:de:bsz:25-freidok-2277314
• Rechteinformation: Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
• Letzte Aktualisierung: 25.03.2025, 13:52 MEZ