title: Macromolecular diffractive imaging using imperfect crystals.
abstract: The three-dimensional structures of macromolecules and their complexes are mainly elucidated by X-ray protein crystallography. A major limitation of this method is access to high-quality crystals, which is necessary to ensure X-ray diffraction extends to sufficiently large scattering angles and hence yields information of sufficiently high resolution with which to solve the crystal structure. The observation that crystals with reduced unit-cell volumes and tighter macromolecular packing often produce higher-resolution Bragg peaks suggests that crystallographic resolution for some macromolecules may be limited not by their heterogeneity, but by a deviation of strict positional ordering of the crystalline lattice. Such displacements of molecules from the ideal lattice give rise to a continuous diffraction pattern that is equal to the incoherent sum of diffraction from rigid individual molecular complexes aligned along several discrete crystallographic orientations and that, consequently, contains more information than Bragg peaks alone. Although such continuous diffraction patterns have long been observed--and are of interest as a source of information about the dynamics of proteins--they have not been used for structure determination. Here we show for crystals of the integral membrane protein complex photosystem II that lattice disorder increases the information content and the resolution of the diffraction pattern well beyond the 4.5-ångström limit of measurable Bragg peaks, which allows us to phase the pattern directly. Using the molecular envelope conventionally determined at 4.5 ångströms as a constraint, we obtain a static image of the photosystem II dimer at a resolution of 3.5 ångströms. This result shows that continuous diffraction can be used to overcome what have long been supposed to be the resolution limits of macromolecular crystallography, using a method that exploits commonly encountered imperfect crystals and enables model-free phasing.
authors: Center for Free-Electron Laser Science, DESY, 22607 Hamburg, Germany.
authors: Kartik
authors: K
authors: Ayyer
authors: Center for Free-Electron Laser Science, DESY, 22607 Hamburg, Germany.
authors: Oleksandr M
authors: OM
authors: Yefanov
authors: Department of Physics, University of Hamburg, 22761 Hamburg, Germany.
authors: Dominik
authors: D
authors: Oberthür
authors: School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.
authors: Shatabdi
authors: S
authors: Roy-Chowdhury
authors: Center for Free-Electron Laser Science, DESY, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, 22761 Hamburg, Germany.
authors: Lorenzo
authors: L
authors: Galli
authors: Center for Free-Electron Laser Science, DESY, 22607 Hamburg, Germany.
authors: Valerio
authors: V
authors: Mariani
authors: School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.
authors: Shibom
authors: S
authors: Basu
authors: School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.
authors: Jesse
authors: J
authors: Coe
authors: School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.
authors: Chelsie E
authors: CE
authors: Conrad
authors: School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.
authors: Raimund
authors: R
authors: Fromme
authors: School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.
authors: Alexander
authors: A
authors: Schaffer
authors: Center for Free-Electron Laser Science, DESY, 22607 Hamburg, Germany.|School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.
authors: Katerina
authors: K
authors: Dörner
authors: Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.|Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors: Daniel
authors: D
authors: James
authors: School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.|Physics Department, University of Wisconsin, Milwaukee, Wisconsin 53211, USA.
authors: Christopher
authors: C
authors: Kupitz
authors: Department of Physics, University of Hamburg, 22761 Hamburg, Germany.
authors: Markus
authors: M
authors: Metz
authors: Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.|Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors: Garrett
authors: G
authors: Nelson
authors: Center for Free-Electron Laser Science, DESY, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, 22761 Hamburg, Germany.
authors: Paulraj Lourdu
authors: PL
authors: Xavier
authors: Center for Free-Electron Laser Science, DESY, 22607 Hamburg, Germany.
authors: Kenneth R
authors: KR
authors: Beyerlein
authors: Physics Department, University of Wisconsin, Milwaukee, Wisconsin 53211, USA.
authors: Marius
authors: M
authors: Schmidt
authors: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, Hellas, GR-70013 Crete, Greece.
authors: Iosifina
authors: I
authors: Sarrou
authors: Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.|Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors: John C H
authors: JC
authors: Spence
authors: Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.|Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors: Uwe
authors: U
authors: Weierstall
authors: Center for Free-Electron Laser Science, DESY, 22607 Hamburg, Germany.
authors: Thomas A
authors: TA
authors: White
authors: School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.
authors: Jay-How
authors: JH
authors: Yang
authors: Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.|Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors: Yun
authors: Y
authors: Zhao
authors: Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC), National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025, USA.
authors: Mengning
authors: M
authors: Liang
authors: Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC), National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025, USA.
authors: Andrew
authors: A
authors: Aquila
authors: Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC), National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025, USA.
authors: Mark S
authors: MS
authors: Hunter
authors: Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC), National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025, USA.
authors: Joseph S
authors: JS
authors: Robinson
authors: Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC), National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025, USA.
authors: Jason E
authors: JE
authors: Koglin
authors: Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC), National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, California 94025, USA.
authors: Sébastien
authors: S
authors: Boutet
authors: School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.|Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA.
authors: Petra
authors: P
authors: Fromme
authors: Center for Free-Electron Laser Science, DESY, 22607 Hamburg, Germany.
authors: Anton
authors: A
authors: Barty
authors: Center for Free-Electron Laser Science, DESY, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, 22761 Hamburg, Germany.|Centre for Ultrafast Imaging, 22607 Hamburg, Germany.
authors: Henry N
authors: HN
authors: Chapman
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D008958:Models, Molecular; D045332:Photosystem II Protein Complex / Q000737:chemistry*
chemical_list: D045332:Photosystem II Protein Complex