title: In situ macromolecular crystallography using microbeams.
abstract: Despite significant progress in high-throughput methods in macromolecular crystallography, the production of diffraction-quality crystals remains a major bottleneck. By recording diffraction in situ from crystals in their crystallization plates at room temperature, a number of problems associated with crystal handling and cryoprotection can be side-stepped. Using a dedicated goniometer installed on the microfocus macromolecular crystallography beamline I24 at Diamond Light Source, crystals have been studied in situ with an intense and flexible microfocus beam, allowing weakly diffracting samples to be assessed without a manual crystal-handling step but with good signal to noise, despite the background scatter from the plate. A number of case studies are reported: the structure solution of bovine enterovirus 2, crystallization screening of membrane proteins and complexes, and structure solution from crystallization hits produced via a high-throughput pipeline. These demonstrate the potential for in situ data collection and structure solution with microbeams.
authors.affiliation: Life Science Division, Diamond Light Source, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE, England.
authors.forename: Danny
authors.initials: D
authors.lastname: Axford
authors.forename: Robin L
authors.initials: RL
authors.lastname: Owen
authors.forename: Jun
authors.initials: J
authors.lastname: Aishima
authors.forename: James
authors.initials: J
authors.lastname: Foadi
authors.forename: Ann W
authors.initials: AW
authors.lastname: Morgan
authors.forename: James I
authors.initials: JI
authors.lastname: Robinson
authors.forename: Joanne E
authors.initials: JE
authors.lastname: Nettleship
authors.forename: Raymond J
authors.initials: RJ
authors.lastname: Owens
authors.forename: Isabel
authors.initials: I
authors.lastname: Moraes
authors.forename: Elizabeth E
authors.initials: EE
authors.lastname: Fry
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Grimes
authors.forename: Karl
authors.initials: K
authors.lastname: Harlos
authors.forename: Abhay
authors.initials: A
authors.lastname: Kotecha
authors.forename: Jingshan
authors.initials: J
authors.lastname: Ren
authors.forename: Geoff
authors.initials: G
authors.lastname: Sutton
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Walter
authors.forename: David I
authors.initials: DI
authors.lastname: Stuart
authors.forename: Gwyndaf
authors.initials: G
authors.lastname: Evans
mesh_terms: D001419:Bacteria / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization / Q000295:instrumentation*; D018360:Crystallography, X-Ray / Q000295:instrumentation*; D004769:Enterovirus Infections / Q000821:virology; D029650:Enterovirus, Bovine / Q000737:chemistry*; D004867:Equipment Design; D046912:Multiprotein Complexes / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D046912:Multiprotein Complexes