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Verfasst von:Schirò, Giorgio [VerfasserIn]   i
 Schlichting, Ilme [VerfasserIn]   i
Titel:Simple and efficient system for photoconverting light-sensitive proteins in serial crystallography experiments
Verf.angabe:Giorgio Schirò, Joyce Woodhouse, Martin Weik, Ilme Schlichting and Robert L. Shoeman
Umfang:8 S.
Fussnoten:Gesehen am 12.09.2018
Titel Quelle:Enthalten in: Journal of applied crystallography
Jahr Quelle:2017
Band/Heft Quelle:50(2017), 3, S. 932-939
ISSN Quelle:1600-5767
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.
DOI:doi:10.1107/S1600576717006264
URL:Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.

Verlag: http://dx.doi.org/10.1107/S1600576717006264
 Verlag: https://onlinelibrary.wiley.com/doi/abs/10.1107/S1600576717006264
 DOI: https://doi.org/10.1107/S1600576717006264
Datenträger:Online-Ressource
Sprache:eng
K10plus-PPN:1580893716
Verknüpfungen:→ Zeitschrift

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