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

Agata Butryn, Philipp S. Simon, Pierre Aller, Philip Hinchliffe, Ramzi N. Massad, Gabriel Leen, Catherine L. Tooke, Isabel Bogacz, In-Sik Kim, Asmit Bhowmick, Aaron S. Brewster, Nicholas E. Devenish, Jürgen Brem, Jos J. A. G. Kamps, Pauline A. Lang, Patrick Rabe, Danny Axford, John H. Beale, Bradley Davy, Ali Ebrahim, Julien Orlans, Selina L. S. Storm, Tiankun Zhou, Shigeki Owada, Rie Tanaka, Kensuke Tono, Gwyndaf Evans, Robin L. Owen, Frances A. Houle, Nicholas K. Sauter, Christopher J. Schofield, James Spencer, Vittal K. Yachandra, Junko Yano, Jan F. Kern () and Allen M. Orville ()
Additional contact information
Agata Butryn: Harwell Science and Innovation Campus
Philipp S. Simon: Lawrence Berkeley National Laboratory
Pierre Aller: Harwell Science and Innovation Campus
Philip Hinchliffe: University Walk
Ramzi N. Massad: Lawrence Berkeley National Laboratory
Gabriel Leen: Rathmacullig West
Catherine L. Tooke: University Walk
Isabel Bogacz: Lawrence Berkeley National Laboratory
In-Sik Kim: Lawrence Berkeley National Laboratory
Asmit Bhowmick: Lawrence Berkeley National Laboratory
Aaron S. Brewster: Lawrence Berkeley National Laboratory
Nicholas E. Devenish: Harwell Science and Innovation Campus
Jürgen Brem: University of Oxford
Jos J. A. G. Kamps: University of Oxford
Pauline A. Lang: University of Oxford
Patrick Rabe: University of Oxford
Danny Axford: Harwell Science and Innovation Campus
John H. Beale: Harwell Science and Innovation Campus
Bradley Davy: Harwell Science and Innovation Campus
Ali Ebrahim: Harwell Science and Innovation Campus
Julien Orlans: Harwell Science and Innovation Campus
Selina L. S. Storm: Harwell Science and Innovation Campus
Tiankun Zhou: Harwell Science and Innovation Campus
Shigeki Owada: RIKEN SPring-8 Center
Rie Tanaka: RIKEN SPring-8 Center
Kensuke Tono: RIKEN SPring-8 Center
Gwyndaf Evans: Harwell Science and Innovation Campus
Robin L. Owen: Harwell Science and Innovation Campus
Frances A. Houle: Lawrence Berkeley National Laboratory
Nicholas K. Sauter: Lawrence Berkeley National Laboratory
Christopher J. Schofield: University of Oxford
James Spencer: University Walk
Vittal K. Yachandra: Lawrence Berkeley National Laboratory
Junko Yano: Lawrence Berkeley National Laboratory
Jan F. Kern: Lawrence Berkeley National Laboratory
Allen M. Orville: Harwell Science and Innovation Campus

Nature Communications, 2021, vol. 12, issue 1, 1-7

Abstract: 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.

Date: 2021
References: Add references at CitEc
Citations: View citations in EconPapers (2)

Downloads: (external link)
https://www.nature.com/articles/s41467-021-24757-7 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-24757-7

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-021-24757-7

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().