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

A method is reported for collecting room-temperature data from protein crystals under vacuum by protecting them with a thin graphene layer. 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.
Authors:
 [1] ;  [2] ; ; ; ;  [1] ;  [2] ;  [1]
  1. Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 0DE (United Kingdom)
  2. Newcastle University, Newcastle upon Tyne NE2 4HH (United Kingdom)
Publication Date:
OSTI Identifier:
22420138
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography; Journal Volume: 71; Journal Issue: Pt 10; Other Information: PMCID: PMC4601369; PMID: 26457431; PUBLISHER-ID: tz5077; OAI: oai:pubmedcentral.nih.gov:4601369; Copyright (c) Warren et al. 2015; 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)
Country of Publication:
Denmark
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; AQUEOUS SOLUTIONS; CRYSTALS; DEHYDRATION; GRAPHENE; LAYERS; LIQUIDS; PROTEINS; SAMPLE PREPARATION; TEMPERATURE RANGE 0273-0400 K; THIN FILMS; X RADIATION