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In vacuo X-ray data collection from graphene-wrapped protein crystals

DOI: 10.1107/S1399004715014194 DOI Help
PMID: 26457431 PMID Help

Authors: Anna J. Warren (Diamond Light Source) , Adam D. Crawshaw (Newcastle University) , Jose Trincao (Diamond Light Source) , Pierre Aller (Diamond Light Source) , Simon Alcock (Diamond Light Source) , Ioana Nistea (Diamond Light Source) , Paula S. Salgado (Newcastle University) , Gwyndaf Evans (Diamond Light Source)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Acta Crystallographica Section D Biological Crystallography , VOL 71 (10) , PAGES 2079 - 2088

State: Published (Approved)
Published: October 2015

Open Access Open Access

Abstract: The measurement of diffraction data from macromolecular crystal samples held in vacuo holds the promise of a very low X-ray background and zero absorption of incident and scattered beams, leading to better data and the potential for accessing very long X-ray wavelengths (>3 Å) for native sulfur phasing. Maintaining the hydration of protein crystals under vacuum is achieved by the use of liquid jets, as with serial data collection at free-electron lasers, or is side-stepped by cryocooling the samples, as implemented at new synchrotron beamlines. Graphene has been shown to protect crystals from dehydration by creating an extremely thin layer that is impermeable to any exchanges with the environment. Furthermore, owing to its hydrophobicity, most of the aqueous solution surrounding the crystal is excluded during sample preparation, thus eliminating most of the background caused by liquid. Here, it is shown that high-quality data can be recorded at room temperature from graphene-wrapped protein crystals in a rough vacuum. Furthermore, it was observed that graphene protects crystals exposed to different relative humidities and a chemically harsh environment

Journal Keywords: Vacuum; Graphene; Room Temperature; Dehydration; Aqueous Solutions; Crystals; Dehydration; Graphene; Layers; Liquids; Proteins; Sample Preparation; Temperature Range 0273-0400 K; Thin Films; X Radiation

Subject Areas: Biology and Bio-materials


Instruments: I04-Macromolecular Crystallography