The Peptidisc, a Simple Method for Stabilizing Membrane Proteins In
TOOLS AND RESOURCES The Peptidisc, a simple method for stabilizing membrane proteins in detergent-free solution Michael Luke Carlson1, John William Young1, Zhiyu Zhao1, Lucien Fabre1, Daniel Jun2,3, Jianing Li4, Jun Li4, Harveer Singh Dhupar1, Irvin Wason1, Allan T Mills1, J Thomas Beatty3, John S Klassen4, Isabelle Rouiller2, Franck Duong1* 1Department of Biochemistry and Molecular Biology, Faculty of Medicine, Life Sciences Institute, University of British Columbia, Vancouver, Canada; 2Department of Anatomy and Cell Biology, McGill University, Montreal, Canada; 3Department of Microbiology and Immunology, University of British Columbia, Vancouver, Canada; 4Glycomics Centre and Department of Chemistry, University of Alberta, Alberta, Canada Abstract 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
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