A Three-Dimensional Model of the Yeast Transmembrane Sensor Wsc1 Obtained by SMA-Based Detergent-Free Purification and Transmission Electron Microscopy
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https://osnadocs.ub.uni-osnabrueck.de/handle/urn:nbn:de:gbv:700-202106105037
https://osnadocs.ub.uni-osnabrueck.de/handle/urn:nbn:de:gbv:700-202106105037
| Titel: | A Three-Dimensional Model of the Yeast Transmembrane Sensor Wsc1 Obtained by SMA-Based Detergent-Free Purification and Transmission Electron Microscopy |
| Autor(en): | Voskoboynikova, Natalia Karlova, Maria Kurre, Rainer Mulkidjanian, Armen Y. Shaitan, Konstantin V. Sokolova, Olga S. Steinhoff, Heinz-Jürgen Heinisch, Jürgen J. |
| ORCID des Autors: | https://orcid.org/0000-0003-4197-4285 https://orcid.org/0000-0003-2317-0144 https://orcid.org/0000-0002-5137-303X https://orcid.org/0000-0003-4678-232X https://orcid.org/0000-0002-5888-0157 |
| Zusammenfassung: | The cell wall sensor Wsc1 belongs to a small family of transmembrane proteins, which are crucial to sustain cell integrity in yeast and other fungi. Wsc1 acts as a mechanosensor of the cell wall integrity (CWI) signal transduction pathway which responds to external stresses. Here we report on the purification of Wsc1 by its trapping in water-soluble polymer-stabilized lipid nanoparticles, obtained with an amphipathic styrene-maleic acid (SMA) copolymer. The latter was employed to transfer tagged sensors from their native yeast membranes into SMA/lipid particles (SMALPs), which allows their purification in a functional state, i.e., avoiding denaturation. The SMALPs composition was characterized by fluorescence correlation spectroscopy, followed by two-dimensional image acquisition from single particle transmission electron microscopy to build a three-dimensional model of the sensor. The latter confirms that Wsc1 consists of a large extracellular domain connected to a smaller intracellular part by a single transmembrane domain, which is embedded within the hydrophobic moiety of the lipid bilayer. The successful extraction of a sensor from the yeast plasma membrane by a detergent-free procedure into a native-like membrane environment provides new prospects for in vitro structural and functional studies of yeast plasma proteins which are likely to be applicable to other fungi, including plant and human pathogens. |
| Bibliografische Angaben: | Voskoboynikova N, Karlova M, Kurre R, Mulkidjanian AY, Shaitan KV, Sokolova OS, Steinhoff H-J, Heinisch JJ.: A Three-Dimensional Model of the Yeast Transmembrane Sensor Wsc1 Obtained by SMA-Based Detergent-Free Purification and Transmission Electron Microscopy. Journal of Fungi. 2021; 7(2):118. |
| URL: | https://osnadocs.ub.uni-osnabrueck.de/handle/urn:nbn:de:gbv:700-202106105037 |
| Schlagworte: | Wsc1; membrane sensor; SMALP; detergent-free extraction; fluorescence correlation spectroscopy; transmission electron microscopy; 3D reconstruction |
| Erscheinungsdatum: | 5-Feb-2021 |
| Lizenzbezeichnung: | Attribution 4.0 International |
| URL der Lizenz: | http://creativecommons.org/licenses/by/4.0/ |
| Publikationstyp: | Einzelbeitrag in einer wissenschaftlichen Zeitschrift [article] |
| Enthalten in den Sammlungen: | FB05 - Hochschulschriften Open-Access-Publikationsfonds |
Dateien zu dieser Ressource:
| Datei | Beschreibung | Größe | Format | |
|---|---|---|---|---|
| jof_Voskoboyniakova_etal_2021.pdf | 2,46 MB | Adobe PDF | jof_Voskoboyniakova_etal_2021.pdf  Öffnen/Anzeigen |
Diese Ressource wurde unter folgender Copyright-Bestimmung veröffentlicht:
Lizenz von Creative Commons
