Mass Photometry of Membrane Proteins
| Autor: | Dzmitry Ashkinadze, Anna Olerinyova, Adar Sonn-Segev, Cédric Eichmann, Radoslaw Bomba, Joseph Gault, Johannes Schimpf, Lucas S. P. Rudden, Roland Riek, Philipp Kukura, Weston B. Struwe, Jason Greenwald, Adrian H. Kopf, J. Antoinette Killian, Thorsten Friedrich, Matteo T. Degiacomi |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
Chemistry Dimer KcsA potassium channel 010402 general chemistry 01 natural sciences 0104 chemical sciences 03 medical and health sciences chemistry.chemical_compound Membrane Membrane protein Amphiphile Biophysics Particle size Integral membrane protein Nanodisc 030304 developmental biology |
| DOI: | 10.1101/2020.02.28.969287 |
| Popis: | 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, characterising 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. |
| Databáze: | OpenAIRE |
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