Date
2021
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 tem poral 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 crys tallography, 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.
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Description
peer-reviewed
Publisher
Nature Publishing Group
Citation
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Electronic & Computer Engineering
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Leen_2021_On_Demand.pdf
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Keywords
crystallograph, enzyme
ULRR Identifiers
https://hdl.handle.net/10344/10448
 https://doi.org/10.34961/3959
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Funding Information
National Institutes of Health (NIH), Biological Sciences Research Council, Royal Society Wolfson
Sustainable Development Goals
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