7YNH

Catalytic intermediate of copper amine oxidase determined by serial femtosecond X-ray crystallography using a single-flow liquid jet system

7YNH の概要

• エントリーDOI: 10.2210/pdb7ynh/pdb
• 分子名称: Phenylethylamine oxidase, SODIUM ION, PHENYLACETALDEHYDE, ... (5 entities in total)
• 機能のキーワード: topaquinone, copper amine oxidase, serial femtosecond x-ray crystallography, oxidoreductase
• 由来する生物種: Arthrobacter globiformis
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 138240.87

構造登録者

Murakawa, T.,Okajima, T. (登録日: 2022-07-31, 公開日: 2022-11-16, 最終更新日: 2024-11-13)

主引用文献

Murakawa, T.,Suzuki, M.,Fukui, K.,Masuda, T.,Sugahara, M.,Tono, K.,Tanaka, T.,Iwata, S.,Nango, E.,Yano, T.,Tanizawa, K.,Okajima, T.
Serial femtosecond X-ray crystallography of an anaerobically formed catalytic intermediate of copper amine oxidase.
Acta Crystallogr D Struct Biol, 78:1428-1438, 2022

PubMed Abstract: The mechanisms by which enzymes promote catalytic reactions efficiently through their structural changes remain to be fully elucidated. Recent progress in serial femtosecond X-ray crystallography (SFX) using X-ray free-electron lasers (XFELs) has made it possible to address these issues. In particular, mix-and-inject serial crystallography (MISC) is promising for the direct observation of structural changes associated with ongoing enzymic reactions. In this study, SFX measurements using a liquid-jet system were performed on microcrystals of bacterial copper amine oxidase anaerobically premixed with a substrate amine solution. The structure determined at 1.94 Å resolution indicated that the peptidyl quinone cofactor is in equilibrium between the aminoresorcinol and semiquinone radical intermediates, which accumulate only under anaerobic single-turnover conditions. These results show that anaerobic conditions were well maintained throughout the liquid-jet SFX measurements, preventing the catalytic intermediates from reacting with dioxygen. These results also provide a necessary framework for performing time-resolved MISC to study enzymic reaction mechanisms under anaerobic conditions.

PubMed: 36458614
DOI: 10.1107/S2059798322010385

実験手法

X-RAY DIFFRACTION (1.94 Å)