title: Stealth carriers for low-resolution structure determination of membrane proteins in solution.
abstract: Structural studies of membrane proteins remain a great experimental challenge. Functional reconstitution into artificial nanoscale bilayer disc carriers that mimic the native bilayer environment allows the handling of membrane proteins in solution. This enables the use of small-angle scattering techniques for fast and reliable structural analysis. The difficulty with this approach is that the carrier discs contribute to the measured scattering intensity in a highly nontrivial fashion, making subsequent data analysis challenging. Here, an elegant solution to circumvent the intrinsic complexity brought about by the presence of the carrier disc is presented. In combination with small-angle neutron scattering (SANS) and the D2O/H2O-based solvent contrast-variation method, it is demonstrated that it is possible to prepare specifically deuterated carriers that become invisible to neutrons in 100% D2O at the length scales relevant to SANS. These `stealth' carrier discs may be used as a general platform for low-resolution structural studies of membrane proteins using well established data-analysis tools originally developed for soluble proteins.
authors: Niels Bohr Institute, University of Copenhagen, Universitetsparken 5, 2100 Copenhagen, Denmark.
authors: Selma
authors: S
authors: Maric
authors: Niels Bohr Institute, University of Copenhagen, Universitetsparken 5, 2100 Copenhagen, Denmark.
authors: Nicholas
authors: N
authors: Skar-Gislinge
authors: Niels Bohr Institute, University of Copenhagen, Universitetsparken 5, 2100 Copenhagen, Denmark.
authors: Søren
authors: S
authors: Midtgaard
authors: CARB Centre, Department of Chemistry, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark.
authors: Mikkel B
authors: MB
authors: Thygesen
authors: Institut für Medizinische Physik und Biophysik, Medizinische Fakultät, Universität Leipzig, Härtelstrasse 16-18, 04107 Leipzig, Germany.
authors: Jürgen
authors: J
authors: Schiller
authors: Forschungszentrum Jülich GmbH, Jülich Centre for Neutron Scattering, TUM FRM-2, Lichtenbergstrasse 1, 85747 Garching, Germany.
authors: Henrich
authors: H
authors: Frielinghaus
authors: Life Sciences Group, Institut Laue-Langevin, 6 Rue Jules Horowitz, 38042 Grenoble, France.
authors: Martine
authors: M
authors: Moulin
authors: Life Sciences Group, Institut Laue-Langevin, 6 Rue Jules Horowitz, 38042 Grenoble, France.
authors: Michael
authors: M
authors: Haertlein
authors: Life Sciences Group, Institut Laue-Langevin, 6 Rue Jules Horowitz, 38042 Grenoble, France.
authors: V Trevor
authors: VT
authors: Forsyth
authors: Center for Membrane Pumps in Cells and Disease, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, Frederiksberg C, Denmark.
authors: Thomas Günther
authors: TG
authors: Pomorski
authors: Niels Bohr Institute, University of Copenhagen, Universitetsparken 5, 2100 Copenhagen, Denmark.
authors: Lise
authors: L
authors: Arleth
mesh_terms: D003903:Deuterium / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008051:Lipid Bilayers / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics; D008567:Membranes, Artificial; D008958:Models, Molecular; D033363:Neutron Diffraction; D009502:Neutrons; D010713:Phosphatidylcholines / Q000737:chemistry*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D053838:Scattering, Small Angle
chemical_list: D008051:Lipid Bilayers; D008565:Membrane Proteins; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D011994:Recombinant Proteins; D003903:Deuterium
keywords: contrast variation; phospholipid bilayer nanodisc; small-angle neutron scattering