Journal
NATURE COMMUNICATIONS
Volume 8, Issue -, Pages -Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41467-017-01702-1
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Funding
- MEXT XFEL
- MEXT KAKENHI [26220807, 17H05896]
- JSPS KAKENHI [15H03841, 15H01055, 15H00965, JP26105012]
- JST-CREST [JPMJCR15P3]
- JST-PRESTO [JPMJPR12L1, JPMJPR14L9]
- JST
- RIKEN
- Grants-in-Aid for Scientific Research [15H01055, 15H03841, 17H03092, 26105012, 15H05806, 26220807, 17H05896, 15K18487, 15H05476, 15H00965] Funding Source: KAKEN
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Time-resolved serial femtosecond crystallography using an X-ray free electron laser (XFEL) in conjunction with a photosensitive caged-compound offers a crystallographic method to track enzymatic reactions. Here we demonstrate the application of this method using fungal NO reductase, a heme-containing enzyme, at room temperature. Twenty milliseconds after caged-NO photolysis, we identify a NO-bound form of the enzyme, which is an initial intermediate with a slightly bent Fe-N-O coordination geometry at a resolution of 2.1 angstrom. The NO geometry is compatible with those analyzed by XFEL-based cryo-crystallography and QM/MM calculations, indicating that we obtain an intact Fe3+-NO coordination structure that is free of X-ray radiation damage. The slightly bent NO geometry is appropriate to prevent immediate NO dissociation and thus accept H- from NADH. The combination of using XFEL and a caged-compound is a powerful tool for determining functional enzyme structures during catalytic reactions at the atomic level.
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