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  Simple and efficient system for photoconverting light-sensitive proteins in serial crystallography experiments

Schirò, G., Woodhouse, J., Weik, M., Schlichting, I., & Shoeman, R. L. (2017). Simple and efficient system for photoconverting light-sensitive proteins in serial crystallography experiments. Journal of Applied Crystallography, 50, 932-939. doi:10.1107/S1600576717006264.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002D-6F6F-A Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002D-6F70-4
Genre: Journal Article

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JApplCryst_50_2017_932.pdf (Any fulltext), 617KB
 
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 Creators:
Schirò, Giorgio, Author
Woodhouse, Joyce, Author
Weik, Martin, Author
Schlichting, Ilme1, Author              
Shoeman, Robert L.1, Author              
Affiliations:
1Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Max Planck Society, ou_1497700              

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Free keywords: serial femtosecond crystallography; light-sensitive proteins; pre-illumination; time resolution; instrumentation
 Abstract: Proteins that change their structure in response to light absorption regulate many functional processes in living cells. Moreover, biotechnological approaches like optogenetics and super-resolution fluorescence microscopy recently triggered the generation of new genetically modified photosensitive proteins. Light-induced structural changes in photosensitive proteins can be studied by time-resolved serial femtosecond crystallography (SFX), an X-ray diffraction technique that allows the determination of macromolecular structures at X-ray free-electron lasers from a large number of nano- to micro-sized crystals. This article describes a simple and efficient system for converting photosensitive proteins into light-induced semi-stationary states by inline laser illumination prior to sample injection with a gas-focused liquid jet and subsequent optical pump–X-ray probe exposure. The simple setup of this device makes it suitable for integration into other liquid injectors (like electro-spinning and electro-kinetic injectors) and potentially also in high-viscosity extruders, provided that embedding microcrystals in viscous media does not alter protein photophysical properties. The functioning of the device is demonstrated with an example of a photoswitchable fluorescent protein pre-illuminated (photoactivated) for time-resolved SFX experiments. The device can be easily adapted for the conversion in time-resolved SFX experiments of other microcrystalline proteins, such as photosystems, phytochromes and rhodopsins.

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Language(s): eng - English
 Dates: 2016-12-132017-04-252017
 Publication Status: Published in print
 Pages: 8
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1107/S1600576717006264
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Title: Journal of Applied Crystallography
  Abbreviation : J. Appl. Cryst.
Source Genre: Journal
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Publ. Info: Oxford, England : Blackwell Publishing on behalf of the International Union of Crystallography
Pages: - Volume / Issue: 50 Sequence Number: - Start / End Page: 932 - 939 Identifier: ISSN: 0021-8898
CoNE: https://pure.mpg.de/cone/journals/resource/954925410812