Serial-femtosecond crystallography reveals how a phytochrome variant couples chromophore and protein structural changes

DOI: 10.1126/sciadv.adp2665

Authors: Luisa Sauthof (Charité-Universitätsmedizin Berlin) , Michal Szczepek (Charité-Universitätsmedizin Berlin) , Andrea Schmidt (Charité-Universitätsmedizin Berlin) , Asmit Bhowmick (Lawrence Berkeley National Laboratory) , Medhanjali Dasgupta (Lawrence Berkeley National Laboratory) , Megan J. Mackintosh (The Hebrew University of Jerusalem) , Sheraz Gul (Lawrence Berkeley National Laboratory) , Franklin D. Fuller (Lawrence Berkeley National Laboratory) , Ruchira Chatterjee (Lawrence Berkeley National Laboratory) , Iris D. Young (Lawrence Berkeley National Laboratory) , Norbert Michael (Technische Universität Berlin) , Nicolas Andreas Heyder (Charité-Universitätsmedizin Berlin) , Brian Bauer (Charité-Universitätsmedizin Berlin) , Anja Koch (Charité-Universitätsmedizin Berlin) , Isabel Bogacz (Lawrence Berkeley National Laboratory) , In-Sik Kim (Lawrence Berkeley National Laboratory) , Philipp S. Simon (Lawrence Berkeley National Laboratory) , Agata Butryn (Diamond Light Source; Research Complex at Harwell) , Pierre Aller (Diamond Light Source; Research Complex at Harwell) , Volha U. Chukhutsina (Imperial College London) , James M. Baxter (Imperial College London) , Christopher D. M. Hutchison (Imperial College London) , Dorothee Liebschner (Lawrence Berkeley National Laboratory) , Billy Poon (Lawrence Berkeley National Laboratory) , Nicholas K. Sauter (Lawrence Berkeley National Laboratory) , Mitchell D. Miller (Rice University) , George N. Phillips (Rice University) , Roberto Alonso-Mori (Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory) , Mark S. Hunter (Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory) , Alexander Batyuk (Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory) , Shigeki Owada (Japan Synchrotron Radiation Research Institute; RIKEN SPring-8 Center) , Kensuke Tono (Japan Synchrotron Radiation Research Institute; RIKEN SPring-8 Center) , Rie Tanaka (RIKEN SPring-8 Center; Kyoto University) , Jasper J. Van Thor (Imperial College London) , Norbert Krauß (Karlsruhe Institute of Technology (KIT)) , Tilman Lamparter (Karlsruhe Institute of Technology (KIT)) , Aaron S. Brewster (Lawrence Berkeley National Laboratory) , Igor Schapiro (The Hebrew University of Jerusalem; Technical University Dortmund; University Alliance Ruhr) , Allen M. Orville (Diamond Light Source; Research Complex at Harwell) , Vittal K. Yachandra (Lawrence Berkeley National Laboratory) , Junko Yano (Lawrence Berkeley National Laboratory) , Peter Hildebrandt (Technische Universität Berlin) , Jan F. Kern (Lawrence Berkeley National Laboratory) , Patrick Scheerer (Charité-Universitätsmedizin Berlin)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Science Advances , VOL 11

State: Published (Approved)
Published: May 2025

Abstract: 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.

Subject Areas: Biology and Bio-materials

Diamond Offline Facilities: XFEL-Hub

Facility: BL14.1, BL14.2, BL14.3 at BESSY II; ID23-1, ID23-2, ID29, ID30A-1, ID30A-3, ID30B at ESRF; MFX at LCLS; SACLA

Added On: 02/06/2025 08:12

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sciadv.adp2665.pdf

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Structural biology Biophysics Life Sciences & Biotech

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