title: 'Hit and run' serial femtosecond crystallography of a membrane kinase in the lipid cubic phase.
abstract: The lipid-based bicontinuous cubic mesophase is a nanoporous membrane mimetic with applications in areas that include medicine, personal care products, foods and the basic sciences. An application of particular note concerns it use as a medium in which to grow crystals of membrane proteins for structure determination by X-ray crystallography. At least two variations of the mesophase exist. One is the highly viscous cubic phase, which has well developed long-range order. The other so-called sponge phase is considerably more fluid and lacks long-range order. The sponge phase has recently been shown to be a convenient vehicle for delivering microcrystals of membrane proteins to an X-ray free-electron laser beam for serial femtosecond crystallography (SFX). Unfortunately, the sponge phase approach calls for large amounts of protein that are not always available in the case of membrane proteins. The cubic phase offers the advantage of requiring significantly less protein for SFX but comes with its own challenges. Here, we describe the physico-chemical bases for these challenges, solutions to them and prospects for future uses of lipidic mesophases in the SFX arena.
authors.affiliation: Membrane Structural and Functional Biology Group, School of Medicine and School of Biochemistry and Immunology, Trinity College Dublin, Dublin 2, Republic of Ireland martin.caffrey@tcd.ie.
authors.forename: Martin
authors.initials: M
authors.lastname: Caffrey
authors.affiliation: Membrane Structural and Functional Biology Group, School of Medicine and School of Biochemistry and Immunology, Trinity College Dublin, Dublin 2, Republic of Ireland.
authors.forename: Dianfan
authors.initials: D
authors.lastname: Li
authors.affiliation: Membrane Structural and Functional Biology Group, School of Medicine and School of Biochemistry and Immunology, Trinity College Dublin, Dublin 2, Republic of Ireland.
authors.forename: Nicole
authors.initials: N
authors.lastname: Howe
authors.affiliation: Membrane Structural and Functional Biology Group, School of Medicine and School of Biochemistry and Immunology, Trinity College Dublin, Dublin 2, Republic of Ireland.
authors.forename: Syed T A
authors.initials: ST
authors.lastname: Shah
mesh_terms: D018360:Crystallography, X-Ray / Q000379:methods*; D019852:Diacylglycerol Kinase / Q000737:chemistry* / Q000648:ultrastructure; D004583:Electrons; D007834:Lasers; D008567:Membranes, Artificial; D011487:Protein Conformation; D013696:Temperature; D013997:Time Factors; D014961:X-Ray Diffraction / Q000379:methods*
chemical_list: D008567:Membranes, Artificial; D019852:Diacylglycerol Kinase
keywords: X-ray free-electron laser; crystal structure; enzyme; membrane protein; mesophase; monoacylglycerol