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Philos Trans R Soc Lond B Biol Sci. 2014 Jul 17;369(1647):20130497. doi: 10.1098/rstb.2013.0497.

In vivo crystallography at X-ray free-electron lasers: the next generation of structural biology?

Author information

1
Photon Factory, High Energy Accelerator Research Organization, 1-1 Oho, Tsukuba, Ibaraki 305-0801, Japan.
2
National Center for Advancing Translational Sciences, National Institutes of Health, 9800 Medical Center Drive, Rockville, MD 20850, USA.
3
Department of Cell and Systems Biology, University of Toronto, Toronto, Canada M5S 3G5.
4
Department of Biochemistry and Molecular Biology, Monash University, Building 76, Clayton, Victoria 3800, Australia.
5
Department of Medicinal Biotechnology, University of Tokushima, 1-78 Sho-machi Tokushima, Tokushima 770-8505, Japan.
6
Deparment of Microbiology and Immunology, School of Medicine, Shimane University, Izumo, Shimane 693-8501, Japan.
7
Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
8
Division of Applied Chemistry, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
9
Japan Synchrotron Radiation Research Institute, Kouto 1-1-1, Sayo, Hyogo 679-5198, Japan.
10
RIKEN SPring-8 Center, Kouto 1-1-1, Sayo, Hyogo 679-5148, Japan.
11
Institute for Stem Cell Biology and Regenerative Medicine, Bellary Road, Bangalore 560065, India.
12
Department of Biology, University of Iowa, Iowa City, IA 52242, USA.
13
RIKEN SPring-8 Center, Kouto 1-1-1, Sayo, Hyogo 679-5148, Japan s.iwata@mfour.med.kyoto-u.ac.jp.
14
Photon Factory, High Energy Accelerator Research Organization, 1-1 Oho, Tsukuba, Ibaraki 305-0801, Japan Center for Free-Electron Laser science, Notkestrasse 85, Building 99, Hamburg 22607, Germany leonard.chavas@desy.de.

Abstract

The serendipitous discovery of the spontaneous growth of protein crystals inside cells has opened the field of crystallography to chemically unmodified samples directly available from their natural environment. On the one hand, through in vivo crystallography, protocols for protein crystal preparation can be highly simplified, although the technique suffers from difficulties in sampling, particularly in the extraction of the crystals from the cells partly due to their small sizes. On the other hand, the extremely intense X-ray pulses emerging from X-ray free-electron laser (XFEL) sources, along with the appearance of serial femtosecond crystallography (SFX) is a milestone for radiation damage-free protein structural studies but requires micrometre-size crystals. The combination of SFX with in vivo crystallography has the potential to boost the applicability of these techniques, eventually bringing the field to the point where in vitro sample manipulations will no longer be required, and direct imaging of the crystals from within the cells will be achievable. To fully appreciate the diverse aspects of sample characterization, handling and analysis, SFX experiments at the Japanese SPring-8 angstrom compact free-electron laser were scheduled on various types of in vivo grown crystals. The first experiments have demonstrated the feasibility of the approach and suggest that future in vivo crystallography applications at XFELs will be another alternative to nano-crystallography.

KEYWORDS:

X-ray free-electron laser; in vivo crystallography; serial femtosecond crystallography

PMID:
24914164
PMCID:
PMC4052873
DOI:
10.1098/rstb.2013.0497
[Indexed for MEDLINE]
Free PMC Article
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