title: Nanodisc Films for Membrane Protein Studies by Neutron Reflection: Effect of the Protein Scaffold Choice.
abstract: Nanodisc films are a promising approach to study the equilibrium conformation of membrane bound proteins in native-like environment. Here we compare nanodisc formation for NADPH-dependent cytochrome P450 oxidoreductase (POR) using two different scaffold proteins, MSP1D1 and MSP1E3D1. Despite the increased stability of POR loaded MSP1E3D1 based nanodiscs in comparison to MSP1D1 based nanodiscs, neutron reflection at the silicon-solution interface showed that POR loaded MSP1E3D1 based nanodisc films had poor surface coverage. This was the case, even when incubation was carried out under conditions that typically gave high coverage for empty nanodiscs. The low surface coverage affects the embedded POR coverage in the nanodisc film and limits the structural information that can be extracted from membrane bound proteins within them. Thus, nanodisc reconstitution on the smaller scaffold proteins is necessary for structural studies of membrane bound proteins in nanodisc films.
authors.affiliation: †Nano-Science Center and Department of Chemistry, Faculty of Science, University of Copenhagen, DK-2200, Copenhagen, Denmark.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bertram
authors.affiliation: #Center of Synthetic Biology "bioSYNergy", University of Copenhagen, 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Denmark.
authors.forename: Tomas
authors.initials: T
authors.lastname: Laursen
authors.affiliation: §Institut Laue Langevin, 71 Avenue des Martyrs - CS 20156, 38042 Grenoble Cedex 9, France.
authors.forename: Robert
authors.initials: R
authors.lastname: Barker
authors.affiliation: #Center of Synthetic Biology "bioSYNergy", University of Copenhagen, 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Denmark.
authors.forename: Krutika
authors.initials: K
authors.lastname: Bavishi
authors.affiliation: ⊥Carlsberg Laboratory, 10 Gamle Carlsberg Vej, DK-1799 Copenhagen V, Denmark.|#Center of Synthetic Biology "bioSYNergy", University of Copenhagen, 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Denmark.
authors.forename: Birger Lindberg
authors.initials: BL
authors.lastname: Møller
authors.affiliation: †Nano-Science Center and Department of Chemistry, Faculty of Science, University of Copenhagen, DK-2200, Copenhagen, Denmark.|#Center of Synthetic Biology "bioSYNergy", University of Copenhagen, 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Denmark.
authors.forename: Marité
authors.initials: M
authors.lastname: Cárdenas
mesh_terms: D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D049329:Nanostructures / Q000737:chemistry*; D033363:Neutron Diffraction
chemical_list: D008565:Membrane Proteins; D003577:Cytochrome P-450 Enzyme System