@article{cd1b864a766f4cfa88528737dc55d70e,
title = "Serial crystallography captures dynamic control of sequential electron and proton transfer events in a flavoenzyme",
abstract = "Flavin coenzymes are universally found in biological redox reactions. DNA photolyases, with their flavin chromophore (FAD), utilize blue light for DNA repair and photoreduction. The latter process involves two single-electron transfers to FAD with an intermittent protonation step to prime the enzyme active for DNA repair. Here we use time-resolved serial femtosecond X-ray crystallography to describe how light-driven electron transfers trigger subsequent nanosecond-to-microsecond entanglement between FAD and its Asn/Arg-Asp redox sensor triad. We found that this key feature within the photolyase-cryptochrome family regulates FAD re-hybridization and protonation. After first electron transfer, the FAD•− isoalloxazine ring twists strongly when the arginine closes in to stabilize the negative charge. Subsequent breakage of the arginine–aspartate salt bridge allows proton transfer from arginine to FAD•−. Our molecular videos demonstrate how the protein environment of redox cofactors organizes multiple electron/proton transfer events in an ordered fashion, which could be applicable to other redox systems such as photosynthesis. [Figure not available: see fulltext.]",
author = "Manuel Maestre-Reyna and Yang, {Cheng Han} and Eriko Nango and Huang, {Wei Cheng} and {Ngurah Putu}, {Eka Putra Gusti} and Wu, {Wen Jin} and Wang, {Po Hsun} and Sophie Franz-Badur and Martin Saft and Emmerich, {Hans Joachim} and Wu, {Hsiang Yi} and Lee, {Cheng Chung} and Huang, {Kai Fa} and Chang, {Yao Kai} and Liao, {Jiahn Haur} and Weng, {Jui Hung} and Wael Gad and Chang, {Chiung Wen} and Pang, {Allan H.} and Michihiro Sugahara and Shigeki Owada and Yuhei Hosokawa and Yasumasa Joti and Ayumi Yamashita and Rie Tanaka and Tomoyuki Tanaka and Fangjia Luo and Kensuke Tono and Hsu, {Kai Cheng} and Stephan Kiontke and Igor Schapiro and Roberta Spadaccini and Antoine Royant and Junpei Yamamoto and So Iwata and Essen, {Lars Oliver} and Yoshitaka Bessho and Tsai, {Ming Daw}",
note = "Funding Information: This work was supported by Academia Sinica and the Taiwan Protein Project funded by the Ministry of Science and Technology (grants nos. AS-KPQ-105-TPP and AS-KPQ-109-TPP2; M.-D.T.), and in part by JSPS KAKENHI (16K01942) to Y.B., by the Air Force Office of Scientific Research (AFOSR; grant no. FA9550-14-1-0409) and the German Research Foundation (DFG, grant no. ES152/18) to L.-O.E. and by Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS) from the Japan Agency for Medical Research and Development (AMED) to S.I. The XFEL experiments were performed at BL2 of SACLA with the approval of the Japan Synchrotron Radiation Research Institute (JASRI) (proposals nos. 2017A8019, 2017B8052, 2018A8008, 2018B8031, 2019A8014 and 2019B8005). We thank T.-C. Hsiao and M.-l. Wu for their assistance with sample preparation, T. Tanaka, T. Arima, Y. Matsuura, H. Naitow, N. Kunishima, T. Kin and the members of the Engineering Support Team of SACLA for help during our X-ray experiments, T. Nakane for the introduction to CrystFEL, all staff members of the TPS05A beamline, NSRRC, a national user facility supported by MOST, R.O.C. (Taiwan) and in particular C.-C. Tseng and C.-K. Chou for help in setting up non-standard conditions for crystal testing, H.-L. Shr and S.-G. Shyu for assisting with crystallization under non-standard conditions, T. C. Terwilliger (Los Alamos National Laboratory) for his kind help in understanding the theoretical underpinnings of Bayesian difference refinement, G. Gotthard for assistance with using the icOS platform of the Grenoble Instruct-ERIC Center (ISBG; UAR 3518 CNRS-CEA-UGA-EMBL) within the Grenoble Partnership for Structural Biology (PSB), supported by FRISBI (ANR-10-INBS-0005-02) and GRAL, financed within the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE-0003), and finally A. M. Reyna for help in setting up the necessary scripts and macros for calculating extrapolated structure factors. Funding Information: This work was supported by Academia Sinica and the Taiwan Protein Project funded by the Ministry of Science and Technology (grants nos. AS-KPQ-105-TPP and AS-KPQ-109-TPP2; M.-D.T.), and in part by JSPS KAKENHI (16K01942) to Y.B., by the Air Force Office of Scientific Research (AFOSR; grant no. FA9550-14-1-0409) and the German Research Foundation (DFG, grant no. ES152/18) to L.-O.E. and by Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS) from the Japan Agency for Medical Research and Development (AMED) to S.I. The XFEL experiments were performed at BL2 of SACLA with the approval of the Japan Synchrotron Radiation Research Institute (JASRI) (proposals nos. 2017A8019, 2017B8052, 2018A8008, 2018B8031, 2019A8014 and 2019B8005). We thank T.-C. Hsiao and M.-l. Wu for their assistance with sample preparation, T. Tanaka, T. Arima, Y. Matsuura, H. Naitow, N. Kunishima, T. Kin and the members of the Engineering Support Team of SACLA for help during our X-ray experiments, T. Nakane for the introduction to CrystFEL, all staff members of the TPS05A beamline, NSRRC, a national user facility supported by MOST, R.O.C. (Taiwan) and in particular C.-C. Tseng and C.-K. Chou for help in setting up non-standard conditions for crystal testing, H.-L. Shr and S.-G. Shyu for assisting with crystallization under non-standard conditions, T. C. Terwilliger (Los Alamos National Laboratory) for his kind help in understanding the theoretical underpinnings of Bayesian difference refinement, G. Gotthard for assistance with using the icOS platform of the Grenoble Instruct-ERIC Center (ISBG; UAR 3518 CNRS-CEA-UGA-EMBL) within the Grenoble Partnership for Structural Biology (PSB), supported by FRISBI (ANR-10-INBS-0005-02) and GRAL, financed within the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE-0003), and finally A. M. Reyna for help in setting up the necessary scripts and macros for calculating extrapolated structure factors. Publisher Copyright: {\textcopyright} 2022, The Author(s), under exclusive licence to Springer Nature Limited.",
year = "2022",
month = jun,
doi = "10.1038/s41557-022-00922-3",
language = "English",
volume = "14",
pages = "677--685",
journal = "Nature Chemistry",
issn = "1755-4330",
publisher = "Nature Publishing Group",
number = "6",
}