Details

Autor(en) / Beteiligte

• Butryn, Agata
• Simon, Philipp S
• Aller, Pierre
• Hinchliffe, Philip
• Massad, Ramzi N
• Leen, Gabriel
• Tooke, Catherine L
• Bogacz, Isabel
• Kim, In-Sik
• Bhowmick, Asmit
• Brewster, Aaron S
• Devenish, Nicholas E
• Brem, Jürgen
• Kamps, Jos J A G
• Lang, Pauline A
• Rabe, Patrick
• Axford, Danny
• Beale, John H
• Davy, Bradley
• Ebrahim, Ali
• Orlans, Julien
• Storm, Selina L S
• Zhou, Tiankun
• Owada, Shigeki
• Tanaka, Rie
• Tono, Kensuke
• Evans, Gwyndaf
• Owen, Robin L
• Houle, Frances A
• Sauter, Nicholas K
• Schofield, Christopher J
• Spencer, James
• Yachandra, Vittal K
• Yano, Junko
• Kern, Jan F
• Orville, Allen M

Titel

An on-demand, drop-on-drop method for studying enzyme catalysis by serial crystallography

Ist Teil von

• Nature communications, 2021-07, Vol.12 (1), p.4461-4461, Article 4461

Ort / Verlag

England: Nature Publishing Group

Erscheinungsjahr

2021

Link zum Volltext

Quelle

Free E-Journal (出版社公開部分のみ）

Beschreibungen/Notizen

• 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.