Time-resolved crystallography captures light-driven DNA repair

• 成果类型: Article
• 署名作者: Christou, Nina-Eleni; Apostolopoulou, Virginia; Melo, Diogo V. M.; Ruppert, Matthias; Fadini, Alisia; Henkel, Alessandra; Sprenger, Janina; Oberthuer, Dominik; Guenther, Sebastian; Pateras, Anastasios; Mashhour, Aida Rahmani; Yefanov, Oleksandr M.; Galchenkova, Marina; Reinke, Patrick Y. A.; Kremling, Viviane; Scheer, T. Emilie S.; Lange, Esther R.; Middendorf, Philipp; Schubert, Robin; De Zitter, Elke; Lumbao-Conradson, Koya; Herrmann, Jonathan; Rahighi, Simin; Kunavar, Ajda; Beale, Emma V.; Beale, John H.; Cirelli, Claudio; Johnson, Philip J. M.; Dworkowski, Florian; Ozerov, Dmitry; Bertrand, Quentin; Wranik, Maximilian; Bacellar, Camila; Bajt, Sasa; Wakatsuki, Soichi; Sellberg, Jonas A.; Huse, Nils; Turk, Dusan; Chapman, Henry N.; Lane, Thomas J.
• 署名单位: Helmholtz Association; Deutsches Elektronen-Synchrotron (DESY); European XFEL; University of Hamburg; Imperial College London; Communaute Universite Grenoble Alpes; Universite Grenoble Alpes (UGA); CEA; Centre National de la Recherche Scientifique (CNRS); Stanford University; United States Department of Energy (DOE); SLAC National Accelerator Laboratory; Stanford University; University of Ljubljana; Swiss Federal Institutes of Technology Domain; Paul Scherrer Institute; Stanford University; United States Department of Energy (DOE); SLAC National Accelerator Laboratory; Royal Institute of Technology; Slovenian Academy of Sciences & Arts (SASA); Jozef Stefan Institute; University of Hamburg; Horiba Ltd.; University of Cambridge; University of Colorado System; University of Colorado Anschutz Medical Campus; Chapman University System; Chapman University; National Institute of Chemistry - Slovenia
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-12068
• DOI: 10.1126/science.adj4270
• 发表日期: 2023-12-01
• 页码: 1015-+
• 关键词: cyclobutane pyrimidine dimer; crystal-structure; photolyase; mechanisms; flavin; pathways; DYNAMICS; STATES

摘要

Photolyase is an enzyme that uses light to catalyze DNA repair. To capture the reaction intermediates involved in the enzyme's catalytic cycle, we conducted a time-resolved crystallography experiment. We found that photolyase traps the excited state of the active cofactor, flavin adenine dinucleotide (FAD), in a highly bent geometry. This excited state performs electron transfer to damaged DNA, inducing repair. We show that the repair reaction, which involves the lysis of two covalent bonds, occurs through a single-bond intermediate. The transformation of the substrate into product crowds the active site and disrupts hydrogen bonds with the enzyme, resulting in stepwise product release, with the 3 ' thymine ejected first, followed by the 5 ' base.