[PDF][PDF] Mix-and-extrude: high-viscosity sample injection towards time-resolved protein crystallography

M Vakili, H Han, C Schmidt, A Wrona… - Journal of Applied …, 2023 - journals.iucr.org
M Vakili, H Han, C Schmidt, A Wrona, M Kloos, I de Diego, K Dörner, T Geng, C Kim
Journal of Applied Crystallography, 2023journals.iucr.org
Time-resolved crystallography enables the visualization of protein molecular motion during
a reaction. Although light is often used to initiate reactions in time-resolved crystallography,
only a small number of proteins can be activated by light. However, many biological
reactions can be triggered by the interaction between proteins and ligands. The sample
delivery method presented here uses a mix-and-extrude approach based on 3D-printed
microchannels in conjunction with a micronozzle. The diffusive mixing enables the study of …
Time-resolved crystallography enables the visualization of protein molecular motion during a reaction. Although light is often used to initiate reactions in time-resolved crystallography, only a small number of proteins can be activated by light. However, many biological reactions can be triggered by the interaction between proteins and ligands. The sample delivery method presented here uses a mix-and-extrude approach based on 3D-printed microchannels in conjunction with a micronozzle. The diffusive mixing enables the study of the dynamics of samples in viscous media. The device design allows mixing of the ligands and protein crystals in 2 to 20 s. The device characterization using a model system (fluorescence quenching of iq-mEmerald proteins by copper ions) demonstrated that ligand and protein crystals, each within lipidic cubic phase, can be mixed efficiently. The potential of this approach for time-resolved membrane protein crystallography to support the development of new drugs is discussed.
International Union of Crystallography
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