Serial-femtosecond crystallography reveals how a phytochrome variant couples chromophore and protein structural changes
The photoreaction and commensurate structural changes of a chromophore within biological photoreceptors elicit conformational transitions of the protein promoting the switch between deactivated and activated states. We investigated how this coupling is achieved in a bacterial phytochrome variant, Agp2-PAiRFP2. Contrary to classical protein crystallography, which only allows probing (cryo-trapped) stable states, we have used time-resolved serial femtosecond x-ray crystallography (tr-SFX) and pump-probe techniques with various illumination and delay times with respect to photoexcitation of the parent Pfr state. Thus, structural data for seven time frames were sorted into groups of molecular events along the reaction coordinate. They range from chromophore isomerization to the formation of Meta-F, the intermediate that precedes the functional relevant secondary structure transition of the tongue. Structural data for the early events were used to calculate the photoisomerization pathway to complement the experimental data. Late events allow identifying the molecular switch that is linked to the intramolecular proton transfer as a prerequisite for the following structural transitions.
Authors
- Aaron S. Brewster (ORCID: https://orcid.org/0000-0002-0908-7822)
- Michal Szczepek (ORCID: https://orcid.org/0000-0001-5079-6893)
- Mitchell D. Miller (ORCID: https://orcid.org/0000-0003-1626-4943)
- Mark S. Hunter (ORCID: https://orcid.org/0000-0002-0110-7075)
- Jan F Kern (ORCID: https://orcid.org/0000-0002-7272-1603)
- Allen Milster Orville (ORCID: https://orcid.org/0000-0002-7803-1777)
- Billy K. Poon (ORCID: https://orcid.org/0000-0001-9633-6067)
- Megan J. Mackintosh (ORCID: https://orcid.org/0000-0002-4913-0848)
- Nicolas Heyder (ORCID: https://orcid.org/0000-0002-8755-5433)
- Medhanjali Dasgupta (ORCID: https://orcid.org/0000-0003-2509-3699)
- Norbert Krauß (ORCID: https://orcid.org/0000-0002-7128-4632)
- Igor Schapiro (ORCID: https://orcid.org/0000-0001-8536-6869)
- C. Hutchison (ORCID: https://orcid.org/0000-0002-2225-2562)
- A. Batyuk (ORCID: https://orcid.org/0000-0002-9393-2880)
- Pierre Aller (ORCID: https://orcid.org/0000-0002-1793-7030)
- Brian Bauer
- Dorothée Liebschner (ORCID: https://orcid.org/0000-0003-3921-3209)
- Andrea Schmidt (ORCID: https://orcid.org/0000-0003-1985-1908)
- Isabel Bogacz (ORCID: https://orcid.org/0000-0003-1493-3867)
- Vittal K. Yachandra (ORCID: https://orcid.org/0000-0002-3983-7858)
- In‐Sik Kim (ORCID: https://orcid.org/0000-0002-0258-6012)
- I.D. Young (ORCID: https://orcid.org/0000-0003-4713-9504)
- Jasper J. van Thor (ORCID: https://orcid.org/0000-0001-6356-3116)
- James M. Baxter (ORCID: https://orcid.org/0000-0002-4641-6760)
- Anja Koch
- Kensuke Tono (ORCID: https://orcid.org/0000-0003-1218-3759)
- Rie Tanaka (ORCID: https://orcid.org/0000-0003-1062-4792)
- Shigeki Owada (ORCID: https://orcid.org/0000-0002-1451-7612)
- Peter Hildebrandt (ORCID: https://orcid.org/0000-0003-1030-5900)
- Norbert Michael (ORCID: https://orcid.org/0000-0002-6722-416X)
- Agata Butryn (ORCID: https://orcid.org/0000-0002-5227-4770)
- G.N. Phillips (ORCID: https://orcid.org/0000-0002-4171-4603)
- Patrick Scheerer (ORCID: https://orcid.org/0000-0001-5028-2075)
- Nicholas K. Sauter (ORCID: https://orcid.org/0000-0003-2786-6552)
- Luisa Sauthof (ORCID: https://orcid.org/0000-0002-6727-8395)
- Roberto Alonso‐Mori (ORCID: https://orcid.org/0000-0002-5357-0934)
- Tilman Lamparter (ORCID: https://orcid.org/0000-0003-4327-9737)
- Philipp S. Simon (ORCID: https://orcid.org/0000-0002-2859-4475)
- Volha U. Chukhutsina (ORCID: https://orcid.org/0000-0003-0304-5573)
- Junko Yano (ORCID: https://orcid.org/0000-0001-6308-9071)
- Asmit Bhowmick (ORCID: https://orcid.org/0000-0002-6154-6635)
- Ruchira Chatterjee (ORCID: https://orcid.org/0000-0002-0865-061X)
- Franklin D. Fuller (ORCID: https://orcid.org/0000-0002-3773-7087)
- Sheraz Gul (ORCID: https://orcid.org/0000-0001-8920-8737)
Institutions
- Karlsruhe Institute of Technology (DE)
- Lawrence Berkeley National Laboratory (US)
- Hebrew University of Jerusalem (IL)
- TU Dortmund University (DE)
- Kyoto University (JP)
- SLAC National Accelerator Laboratory (US)
- Diamond Light Source (GB)
- Humboldt-Universität zu Berlin (DE)
- Research Complex at Harwell (GB)
- SPring-8 (JP)
- Japan Synchrotron Radiation Research Institute (JP)
- Linac Coherent Light Source (US)
- Technische Universität Berlin (DE)
- Imperial College London (GB)
- Rice University (US)
- Ruhr University Bochum (DE)
Publication Details
- Journal
- Science Advances
- Published
- 2025-05-28
- DOI
- https://doi.org/10.1126/sciadv.adp2665
- Citations
- 4
- Primary Topic
- Light effects on plants
- Type
- article
- Field-Weighted Citation Impact
- 7.00