7BHM

Crystal structure of hen egg white lysozyme using drop-on-drop SFX method - 0.7 s mixing with N-acetyl-D-glucosamine.

7BHM の概要

• エントリーDOI: 10.2210/pdb7bhm/pdb
• 関連するPDBエントリー: 7BHK 7BHL 7BHN
• 分子名称: Lysozyme, 2-acetamido-2-deoxy-alpha-D-glucopyranose, CHLORIDE ION, ... (5 entities in total)
• 機能のキーワード: lysozyme, sfx, xfel, 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: 14717.17

構造登録者

Butryn, A.,Orville, A.M. (登録日: 2021-01-11, 公開日: 2021-07-21, 最終更新日: 2024-10-23)

主引用文献

Butryn, A.,Simon, P.S.,Aller, P.,Hinchliffe, P.,Massad, R.N.,Leen, G.,Tooke, C.L.,Bogacz, I.,Kim, I.S.,Bhowmick, A.,Brewster, A.S.,Devenish, N.E.,Brem, J.,Kamps, J.J.A.G.,Lang, P.A.,Rabe, P.,Axford, D.,Beale, J.H.,Davy, B.,Ebrahim, A.,Orlans, J.,Storm, S.L.S.,Zhou, T.,Owada, S.,Tanaka, R.,Tono, K.,Evans, G.,Owen, R.L.,Houle, F.A.,Sauter, N.K.,Schofield, C.J.,Spencer, J.,Yachandra, V.K.,Yano, J.,Kern, J.F.,Orville, A.M.
An on-demand, drop-on-drop method for studying enzyme catalysis by serial crystallography.
Nat Commun, 12:4461-4461, 2021

PubMed Abstract: Serial femtosecond crystallography has opened up many new opportunities in structural biology. In recent years, several approaches employing light-inducible systems have emerged to enable time-resolved experiments that reveal protein dynamics at high atomic and temporal resolutions. However, very few enzymes are light-dependent, whereas macromolecules requiring ligand diffusion into an active site are ubiquitous. In this work we present a drop-on-drop sample delivery system that enables the study of enzyme-catalyzed reactions in microcrystal slurries. The system delivers ligand solutions in bursts of multiple picoliter-sized drops on top of a larger crystal-containing drop inducing turbulent mixing and transports the mixture to the X-ray interaction region with temporal resolution. We demonstrate mixing using fluorescent dyes, numerical simulations and time-resolved serial femtosecond crystallography, which show rapid ligand diffusion through microdroplets. The drop-on-drop method has the potential to be widely applicable to serial crystallography studies, particularly of enzyme reactions with small molecule substrates.

PubMed: 34294694
DOI: 10.1038/s41467-021-24757-7

実験手法

X-RAY DIFFRACTION (1.45 Å)