The Peptidisc, a simple method for stabilizing membrane proteins in detergent-free solution
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- Michael Luke Carlson
- Department of Biochemistry and Molecular Biology, Faculty of Medicine, Life Sciences Institute, University of British Columbia, Vancouver, Canada
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- John William Young
- Department of Biochemistry and Molecular Biology, Faculty of Medicine, Life Sciences Institute, University of British Columbia, Vancouver, Canada
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- Zhiyu Zhao
- Department of Biochemistry and Molecular Biology, Faculty of Medicine, Life Sciences Institute, University of British Columbia, Vancouver, Canada
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- Lucien Fabre
- Department of Biochemistry and Molecular Biology, Faculty of Medicine, Life Sciences Institute, University of British Columbia, Vancouver, Canada
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- Daniel Jun
- Department of Anatomy and Cell Biology, McGill University, Montreal, Canada
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- Jianing Li
- Glycomics Centre and Department of Chemistry, University of Alberta, Alberta, Canada
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- Jun Li
- Glycomics Centre and Department of Chemistry, University of Alberta, Alberta, Canada
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- Harveer Singh Dhupar
- Department of Biochemistry and Molecular Biology, Faculty of Medicine, Life Sciences Institute, University of British Columbia, Vancouver, Canada
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- Irvin Wason
- Department of Biochemistry and Molecular Biology, Faculty of Medicine, Life Sciences Institute, University of British Columbia, Vancouver, Canada
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- Allan T Mills
- Department of Biochemistry and Molecular Biology, Faculty of Medicine, Life Sciences Institute, University of British Columbia, Vancouver, Canada
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- J Thomas Beatty
- Department of Microbiology and Immunology, University of British Columbia, Vancouver, Canada
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- John S Klassen
- Glycomics Centre and Department of Chemistry, University of Alberta, Alberta, Canada
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- Isabelle Rouiller
- Department of Anatomy and Cell Biology, McGill University, Montreal, Canada
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- Franck Duong
- Department of Biochemistry and Molecular Biology, Faculty of Medicine, Life Sciences Institute, University of British Columbia, Vancouver, Canada
書誌事項
- 公開日
- 2018-08-15
- 権利情報
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- http://creativecommons.org/licenses/by/4.0/
- http://creativecommons.org/licenses/by/4.0/
- http://creativecommons.org/licenses/by/4.0/
- DOI
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- 10.7554/elife.34085
- 公開者
- eLife Sciences Publications, Ltd
説明
<jats:p>Membrane proteins are difficult to work with due to their insolubility in aqueous solution and quite often their poor stability in detergent micelles. Here, we present the peptidisc for their facile capture into water-soluble particles. Unlike the nanodisc, which requires scaffold proteins of different lengths and precise amounts of matching lipids, reconstitution of detergent solubilized proteins in peptidisc only requires a short amphipathic bi-helical peptide (NSPr) and no extra lipids. Multiple copies of the peptide wrap around to shield the membrane-exposed part of the target protein. We demonstrate the effectiveness of this ‘one size fits all’ method using five different membrane protein assemblies (MalFGK2, FhuA, SecYEG, OmpF, BRC) during ‘on-column’, ‘in-gel’, and ‘on-bead’ reconstitution embedded within the membrane protein purification protocol. The peptidisc method is rapid and cost-effective, and it may emerge as a universal tool for high-throughput stabilization of membrane proteins to advance modern biological studies.</jats:p>
収録刊行物
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- eLife
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eLife 7 e34085-, 2018-08-15
eLife Sciences Publications, Ltd
