title: Two-dimensional crystallization of integral membrane proteins for electron crystallography.
abstract: Although membrane proteins make up 30% of the proteome and are a common target for therapeutic drugs, determination of their atomic structure remains a technical challenge. Electron crystallography represents an alternative to the conventional methods of X-ray diffraction and NMR and relies on the formation of two-dimensional crystals. These crystals are produced by reconstituting purified, detergent-solubilized membrane proteins back into the native environment of a lipid bilayer. This chapter reviews methods for producing two-dimensional crystals and for screening them by negative stain electron microscopy. In addition, we show examples of the different morphologies that are commonly obtained and describe basic image analysis procedures that can be used to evaluate their promise for structure determination by cryoelectron microscopy.
authors.affiliation: Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY, USA. stokes@saturn.med.nyu.edu
authors.forename: David L
authors.initials: DL
authors.lastname: Stokes
authors.forename: William J
authors.initials: WJ
authors.lastname: Rice
authors.forename: Minghui
authors.initials: M
authors.lastname: Hu
authors.forename: Changki
authors.initials: C
authors.lastname: Kim
authors.forename: Iban
authors.initials: I
authors.lastname: Ubarretxena-Belandia
mesh_terms: D020285:Cryoelectron Microscopy; D003461:Crystallography / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry* / Q000648:ultrastructure; D008854:Microscopy, Electron
chemical_list: D008565:Membrane Proteins