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Phytochromes are a family of photoreceptors that control light responses of plants, fungi and bacteria. A sequence of structural changes, which is not yet fully understood, leads to activation of an output domain. Time-resolved serial femtosecond crystallography (SFX) can potentially shine light on these conformational changes. Here we report the room temperature crystal structure of the chromophore-binding domains of the Deinococcus radiodurans phytochrome at 2.1 Å resolution. The structure was obtained by serial femtosecond X-ray crystallography from microcrystals at an X-ray free electron laser. We find overall good agreement compared to a crystal structure at 1.35 Å resolution derived from conventional crystallography at cryogenic temperatures, which we also report here. The thioether linkage between chromophore and protein is subject to positional ambiguity at the synchrotron, but is fully resolved with SFX. The study paves the way for time-resolved structural investigations of the phytochrome photocycle with time-resolved SFX.
ContributorsEdlund, Petra (Author) / Takala, Heikki (Author) / Claesson, Elin (Author) / Henry, Leocadie (Author) / Dods, Robert (Author) / Lehtivuori, Heli (Author) / Panman, Matthijs (Author) / Pande, Kanupriya (Author) / White, Thomas (Author) / Nakane, Takanori (Author) / Berntsson, Oskar (Author) / Gustavsson, Emil (Author) / Bath, Petra (Author) / Modi, Vaibhav (Author) / Roy Chowdhury, Shatabdi (Author) / Zook, James (Author) / Berntsen, Peter (Author) / Pandey, Suraj (Author) / Poudyal, Ishwor (Author) / Tenboer, Jason (Author) / Kupitz, Christopher (Author) / Barty, Anton (Author) / Fromme, Petra (Author) / Koralek, Jake D. (Author) / Tanaka, Tomoyuki (Author) / Spence, John (Author) / Liang, Mengning (Author) / Hunter, Mark S. (Author) / Boutet, Sebastien (Author) / Nango, Eriko (Author) / Moffat, Keith (Author) / Groenhof, Gerrit (Author) / Ihalainen, Janne (Author) / Stojkovic, Emina A. (Author) / Schmidt, Marius (Author) / Westenhoff, Sebastian (Author) / College of Liberal Arts and Sciences (Contributor) / School of Molecular Sciences (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / Department of Physics (Contributor)
Created2016-10-19
ContributorsMarine Biological Laboratory (Woods Hole, Mass.) (Publisher) / Arizona Board of Regents (Publisher)
Created1939
ContributorsMarine Biological Laboratory (Woods Hole, Mass.) (Creator) / Marine Biological Laboratory (Publisher)
Created2006
ContributorsMarine Biological Laboratory (Woods Hole, Mass.) (Creator) / Marine Biological Laboratory (Publisher)
Created2007
ContributorsMarine Biological Laboratory (Woods Hole, Mass.) (Publisher) / Arizona Board of Regents (Publisher)
Created1944
ContributorsMarine Biological Laboratory (Woods Hole, Mass.) (Publisher) / Arizona Board of Regents (Publisher)
Created1945
ContributorsMarine Biological Laboratory (Woods Hole, Mass.) (Publisher) / Arizona Board of Regents (Publisher, Publisher)
Created1948
ContributorsMarine Biological Laboratory (Woods Hole, Mass.) (Creator) / Marine Biological Laboratory (Publisher)
Created1996
ContributorsMarine Biological Laboratory (Woods Hole, Mass.) (Publisher) / Arizona Board of Regents (Publisher)
Created1955
ContributorsMarine Biological Laboratory (Woods Hole, Mass.) (Publisher, Publisher) / Arizona Board of Regents (Publisher, Publisher)
Created1960