Endoplasmic reticulum phospholipid scramblase activity revealed after protein reconstitution into giant unilamellar vesicles containing a photostable lipid reporter
- Transbilayer movement of phospholipids in biological membranes is mediated by a diverse set of lipid transporters. Among them are scramblases that facilitate a rapid bi-directional movement of lipids without metabolic energy input. Here, we established a new fluorescence microscopy-based assay for detecting phospholipid scramblase activity of membrane proteins upon their reconstitution into giant unilamellar vesicles formed from proteoliposomes by electroformation. The assay is based on chemical bleaching of fluorescence of a photostable ATTO-dye labeled phospholipid with the membrane-impermeant reductant sodium dithionite. We demonstrate that this new methodology is suitable for the study of the scramblase activity of the yeast endoplasmic reticulum at single vesicle level.
| Author: | Patricia P.M. MathiassenGND, Anant K. MenonGND, Thomas Günther-PomorskiORCiDGND |
|---|---|
| URN: | urn:nbn:de:hbz:294-90759 |
| DOI: | https://doi.org/10.1038/s41598-021-93664-0 |
| Parent Title (English): | Scientific reports |
| Publisher: | Springer Nature |
| Place of publication: | Berlin |
| Document Type: | Article |
| Language: | English |
| Date of Publication (online): | 2022/06/27 |
| Date of first Publication: | 2021/07/13 |
| Publishing Institution: | Ruhr-Universität Bochum, Universitätsbibliothek |
| Tag: | Open Access Fonds |
| Volume: | 11 |
| Issue: | Artikel 14364 |
| First Page: | 14364-1 |
| Last Page: | 14364-14 |
| Note: | Article Processing Charge funded by the Open Access Publication Fund of Ruhr-Universität Bochum. |
| Institutes/Facilities: | Lehrstuhl Biochemie II, Molekulare Biochemie |
| Dewey Decimal Classification: | Naturwissenschaften und Mathematik / Biowissenschaften, Biologie, Biochemie |
| faculties: | Fakultät für Chemie und Biochemie |
| Licence (English): |  Creative Commons - CC BY 4.0 - Attribution 4.0 International |
