title: A lipidic-sponge phase screen for membrane protein crystallization.
abstract: A major current deficit in structural biology is the lack of high-resolution structures of eukaryotic membrane proteins, many of which are key drug targets for the treatment of disease. Numerous eukaryotic membrane proteins require specific lipids for their stability and activity, and efforts to crystallize and solve the structures of membrane proteins that do not address the issue of lipids frequently end in failure rather than success. To help address this problem, we have developed a sparse matrix crystallization screen consisting of 48 lipidic-sponge phase conditions. Sponge phases form liquid lipid bilayer environments which are suitable for conventional hanging- and sitting-drop crystallization experiments. Using the sponge phase screen, we obtained crystals of several different membrane proteins from bacterial and eukaryotic sources. We also demonstrate how the screen may be manipulated by incorporating specific lipids such as cholesterol; this modification led to crystals being recovered from a bacterial photosynthetic core complex.
authors.affiliation: Department of Chemical and Biological Engineering, Molecular Biotechnology, Chalmers University of Technology, SE-405 30 Gothenburg, Sweden.
authors.forename: Annemarie B
authors.initials: AB
authors.lastname: Wöhri
authors.forename: Linda C
authors.initials: LC
authors.lastname: Johansson
authors.forename: Pia
authors.initials: P
authors.lastname: Wadsten-Hindrichsen
authors.forename: Weixiao Y
authors.initials: WY
authors.lastname: Wahlgren
authors.forename: Gerhard
authors.initials: G
authors.lastname: Fischer
authors.forename: Rob
authors.initials: R
authors.lastname: Horsefield
authors.forename: Gergely
authors.initials: G
authors.lastname: Katona
authors.forename: Maria
authors.initials: M
authors.lastname: Nyblom
authors.forename: Fredrik
authors.initials: F
authors.lastname: Oberg
authors.forename: Gillian
authors.initials: G
authors.lastname: Young
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Cogdell
authors.forename: Niall J
authors.initials: NJ
authors.lastname: Fraser
authors.forename: Sven
authors.initials: S
authors.lastname: Engström
authors.forename: Richard
authors.initials: R
authors.lastname: Neutze
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D003460:Crystallization / Q000379:methods*; D003902:Detergents / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry / Q000648:ultrastructure*; D053838:Scattering, Small Angle; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D003902:Detergents; D008051:Lipid Bilayers; D008565:Membrane Proteins