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Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers
Published on: February 28, 2019
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Mix-and-extrude: high-viscosity sample injection towards time-resolved protein crystallography
Mohammad Vakili1, Huijong Han1, Christina Schmidt1
1European XFEL GmbH, Holzkoppel 4, Schenefeld 22869, Germany.
Journal of Applied Crystallography
|August 9, 2023
Summary
A new method uses 3D-printed microchannels for rapid mixing of protein crystals and ligands. This technique advances time-resolved crystallography for studying protein dynamics and drug development.
Area of Science:
- Structural Biology
- Biochemistry
- Crystallography
Background:
- Time-resolved crystallography visualizes protein dynamics during reactions.
- Current methods often rely on light activation, limiting applicability.
- Many biological reactions are ligand-triggered.
Purpose of the Study:
- To develop a novel sample delivery method for time-resolved crystallography.
- To enable the study of ligand-triggered protein reactions.
- To support drug development through enhanced membrane protein studies.
Main Methods:
- A mix-and-extrude approach utilizing 3D-printed microchannels and a micronozzle.
- Diffusive mixing in viscous media for studying sample dynamics.
- Efficient mixing of ligands and protein crystals within lipidic cubic phase in 2-20 seconds.
Main Results:
- Successful characterization using a fluorescence quenching model system (iq-mEmerald proteins and copper ions).
- Demonstrated efficient mixing of ligand and protein crystals.
- Validated the method's capability for studying dynamics in viscous media.
Conclusions:
- The developed method facilitates ligand-triggered reactions for time-resolved crystallography.
- This approach enhances the study of membrane protein dynamics.
- Potential applications in accelerating new drug development.
