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Visualization of membrane protein crystals in lipid cubic phase using X-ray imaging

DOI: 10.1107/S0907444913011359 DOI Help
PMID: 23793151 PMID Help

Authors: Wes Armour (Diamond Light Source) , Danny Axford (Diamond Light Source) , Mark Basham (Diamond Light Source) , Thomas Connolley (Diamond Light Source) , David R. Hall (Diamond Light Source) , Sam Horrell (Diamond Light Source; University of Liverpool) , Katherine Mcauley (Diamond Light Source) , Vitaliy Mykhaylyk (Diamond Light Source) , Armin Wagner (Diamond Light Source) , Gwyndaf Evans (Diamond Light Source) , Anna Warren (Diamond Light Source)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Acta Crystallographica Section D Biological Crystallography , VOL 69 , PAGES 1252 - 1259

State: Published (Approved)
Published: July 2013

Open Access Open Access

Abstract: The focus in macromolecular crystallography is moving towards even more challenging target proteins that often crystallize on much smaller scales and are frequently mounted in opaque or highly refractive materials. It is therefore essential that X-ray beamline technology develops in parallel to accommodate such difficult samples. In this paper, the use of X-ray microradiography and microtomography is reported as a tool for crystal visualization, location and characterization on the macromolecular crystallography beamlines at the Diamond Light Source. The technique is particularly useful for microcrystals and for crystals mounted in opaque materials such as lipid cubic phase. X-ray diffraction raster scanning can be used in combination with radiography to allow informed decision-making at the beamline prior to diffraction data collection. It is demonstrated that the X-ray dose required for a full tomography measurement is similar to that for a diffraction grid-scan, but for sample location and shape estimation alone just a few radiographic projections may be required.

Journal Keywords: Crystals; Diamonds; Grids; Light Sources; Materials; Membrane Proteins; Shape; Tomography; X-Ray Diffraction

Subject Areas: Technique Development, Biology and Bio-materials

Diamond Offline Facilities: Membrane Protein Laboratory (MPL)
Instruments: I03-Macromolecular Crystallography , I04-Macromolecular Crystallography