7WKR

Room temperature structure of lysozyme solved by serial synchrotron crystallography

7WKR の概要

• エントリーDOI: 10.2210/pdb7wkr/pdb
• 分子名称: Lysozyme C, CHLORIDE ION (3 entities in total)
• 機能のキーワード: lysozyme, serial crystallography, room temperature, synchrotron, hydrolase
• 由来する生物種: Gallus gallus (chicken)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 14402.07

構造登録者

Nam, K.H. (登録日: 2022-01-11, 公開日: 2022-02-02, 最終更新日: 2024-10-23)

主引用文献

Lee, K.,Kim, J.,Baek, S.,Park, J.,Park, S.,Lee, J.L.,Chung, W.K.,Cho, Y.,Nam, K.H.
Combination of an inject-and-transfer system for serial femtosecond crystallography.
J.Appl.Crystallogr., 55:813-822, 2022

PubMed Abstract: Serial femtosecond crystallography (SFX) enables the determination of room-temperature crystal structures of macromolecules with minimized radiation damage and provides time-resolved molecular dynamics by pump-probe or mix-and-inject experiments. In SFX, a variety of sample delivery methods with unique advantages have been developed and applied. The combination of existing sample delivery methods can enable a new approach to SFX data collection that combines the advantages of the individual methods. This study introduces a combined inject-and-transfer system (BITS) method for sample delivery in SFX experiments: a hybrid injection and fixed-target scanning method. BITS allows for solution samples to be reliably deposited on ultraviolet ozone (UVO)-treated polyimide films, at a minimum flow rate of 0.5 nl min, in both vertical and horizontal scanning modes. To utilize BITS in SFX experiments, lysozyme crystal samples were embedded in a viscous lard medium and injected at flow rates of 50-100 nl min through a syringe needle onto a UVO-treated polyimide film, which was mounted on a fixed-target scan stage. The crystal samples deposited on the film were raster scanned with an X-ray free electron laser using a motion stage in both horizontal and vertical directions. Using the BITS method, the room-temperature structure of lysozyme was successfully determined at a resolution of 2.1 Å, and thus BITS could be utilized in future SFX experiments.

PubMed: 35979068
DOI: 10.1107/S1600576722005556

実験手法

X-RAY DIFFRACTION (1.6 Å)