title: Reconstitution of Na(+),K(+)-ATPase in Nanodiscs.
abstract: Nanodiscs are disc-shaped self-assembled lipid bilayers encircled by membrane scaffolding proteins derived from Apolipoprotein A-1 (apo A-1). They constitute a versatile tool for studying membrane proteins since reconstitution into nanodiscs allows studies of the membrane proteins in detergent-free aqueous solutions in a lipid bilayer. Here, we apply the technique to the Na(+),K(+)-ATPase (NKA) from pig kidney using Membrane Scaffolding Protein 1 D1 (MSP1D1). Contrary to other reports, the nanodiscs obtained by our protocol are built up of the native lipids originally present in the detergent solubilized sample together with the NKA.
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10C, 8000, Aarhus C, Denmark.|Centre for Membrane Pumps in Cells and Disease - PUMPkin, Danish National Research Foundation, Gustav Wieds Vej 10C, 8000, Aarhus C, Denmark.
authors.forename: Jonas Lindholt
authors.initials: JL
authors.lastname: Gregersen
authors.affiliation: Centre for Membrane Pumps in Cells and Disease - PUMPkin, Danish National Research Foundation, Gustav Wieds Vej 10C, 8000, Aarhus C, Denmark.|Department of Biomedicine, Aarhus University, Ole Worms Allé 6, bldg. 1185, 8000, Aarhus C, Denmark.
authors.forename: Natalya U
authors.initials: NU
authors.lastname: Fedosova
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10C, 8000, Aarhus C, Denmark.
authors.forename: Poul
authors.initials: P
authors.lastname: Nissen
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10C, 8000, Aarhus C, Denmark. thb@mbg.au.dk.|Centre for Membrane Pumps in Cells and Disease - PUMPkin, Danish National Research Foundation, Gustav Wieds Vej 10C, 8000, Aarhus C, Denmark. thb@mbg.au.dk.|Danish Research Institute for Translational Neuroscience - DANDRITE, Nordic EMBL Partnership of Molecular Medicine, Gustav Wieds Vej 10C, 8000, Aarhus C, Denmark. thb@mbg.au.dk.
authors.forename: Thomas
authors.initials: T
authors.lastname: Boesen
mesh_terms: D000818:Animals; D016632:Apolipoprotein A-I / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods*; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry / Q000378:metabolism*; D012995:Solubility; D013552:Swine
chemical_list: D016632:Apolipoprotein A-I; D008051:Lipid Bilayers; D000254:Sodium-Potassium-Exchanging ATPase
keywords: Na+,K+-ATPase; Nanodisc; Native lipids; Scaffolding protein; Self-assembled lipid bilayer; Self-assembly