In situ macromolecular crystallography using microbeams

Journal Article · Tue Apr 17 00:00:00 EDT 2012 · Acta Crystallographica. Section D: Biological Crystallography

DOI:https://doi.org/10.1107/S0907444912006749· OSTI ID:22515200

Axford, Danny; Owen, Robin L.; Aishima, Jun ^[1]; Foadi, James ^[1]; Morgan, Ann W.; Robinson, James I. ^[2]; Nettleship, Joanne E.; Owens, Raymond J. ^[3]; Moraes, Isabel ^[4]; Fry, Elizabeth E.; Grimes, Jonathan M.; Harlos, Karl; Kotecha, Abhay; Ren, Jingshan; Sutton, Geoff; Walter, Thomas S. ^[5]; Stuart, David I. ^[1]; Evans, Gwyndaf ^[1]

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1. Diamond Light Source, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE (United Kingdom)
2. University of Leeds, Leeds LS9 7FT (United Kingdom)
3. Research Complex at Harwell, Rutherford Appleton Laboratory R92, Didcot, Oxfordshire OX11 0DE (United Kingdom)
4. Imperial College, London SW7 2AZ (United Kingdom)
5. University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom)

A sample environment for mounting crystallization trays has been developed on the microfocus beamline I24 at Diamond Light Source. The technical developments and several case studies are described. 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.

• OSTI ID: 22515200
• Journal Information: Acta Crystallographica. Section D: Biological Crystallography, Vol. 68, Issue Pt 5; Other Information: PMCID: PMC4791750; PMID: 22525757; PUBLISHER-ID: tz5002; OAI: oai:pubmedcentral.nih.gov:4791750; Copyright (c) Axford et al. 2012; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA); ISSN 0907-4449
• Country of Publication: Denmark
• Language: English

Related Subjects

75 CONDENSED MATTER PHYSICS
SUPERCONDUCTIVITY AND SUPERFLUIDITY
CRYSTALLIZATION
CRYSTALS
DIFFRACTION
LIGHT SOURCES
MATHEMATICAL SOLUTIONS
SYNCHROTRON RADIATION
TEMPERATURE RANGE 0273-0400 K