The Peptidisc, a simple method for stabilizing membrane proteins in detergent-free solution.
Michael Carlson,John W. Young,Zhiyu Zhao,Lucien Fabre,Daniel Jun,Daniel Jun,Jianing Li,Jun Li,Harveer Singh Dhupar,Irvin Wason,Allan Mills,J. Thomas Beatty,John S. Klassen,Isabelle Rouiller,Franck Duong +14 more
TLDR
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.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 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.read more
Citations
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Structural insights into the lipid and ligand regulation of serotonin receptors
Peiyu Xu,Peiyu Xu,Sijie Huang,Sijie Huang,Huibing Zhang,Chunyou Mao,X. Edward Zhou,Xi Cheng,Ícaro Ariel Simon,Dan-Dan Shen,Hsin-Yung Yen,Carol V. Robinson,Kasper Harpsøe,Bo Svensson,Jia Guo,Hualiang Jiang,Hualiang Jiang,David E. Gloriam,Karsten Melcher,Yi Jiang,Yan Zhang,H. Eric Xu,H. Eric Xu +22 more
TL;DR: In this paper, the structural basis for the lipid regulation and basal activation of 5-HT receptors and the panagonism of 5HT remains unclear, and the role of lipids and water molecules in regulating these receptors and their structural basis of ligand recognition is addressed.
Journal ArticleDOI
Megabodies expand the nanobody toolkit for protein structure determination by single-particle cryo-EM
Tomasz Uchański,S. Masiulis,Baptiste Fischer,Valentina Kalichuk,Uriel López-Sánchez,Eleftherios Zarkadas,Miriam Weckener,Andrija Sente,Philip N. Ward,Alexandre Wohlkonig,Thomas Zögg,Han Remaut,James H. Naismith,James H. Naismith,Hugues Nury,Wim F. Vranken,A. Radu Aricescu,A. Radu Aricescu,Els Pardon,Jan Steyaert +19 more
TL;DR: In this paper, the authors demonstrate that engineered nanobodies can also help overcome two major obstacles that limit the resolution of single-particle cryo-electron microscopy reconstructions: particle size and preferential orientation at the water-air interfaces.
Journal ArticleDOI
Nanodiscs: A toolkit for membrane protein science.
TL;DR: Nanodiscs provide a nanometer scale vehicle for the in vivo delivery of amphipathic drugs, therapeutic lipids, tethered nucleic acids, imaging agents and active protein complexes and have found great utility in the study of cellular signaling complexes that assemble on a membrane surface.
Journal ArticleDOI
Native Mass Spectrometry of Membrane Proteins.
Journal ArticleDOI
Profiling the Escherichia coli membrane protein interactome captured in Peptidisc libraries
Michael Carlson,R. Greg Stacey,John W. Young,Irvinder Singh Wason,Zhiyu Zhao,David G Rattray,Nichollas E. Scott,Craig H. Kerr,Mohan Babu,Leonard J. Foster,Franck Duong Van Hoa +10 more
TL;DR: The peptidisc workflow applied to the proteomic field is emerging as a promising novel approach to characterize membrane protein interactions under native expression conditions and without genetic manipulation.
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