Rapid and efficient ambient temperature X-ray crystal structure determination at Turkish Light Source.

Area of Science:

• Structural Biology
• Biophysics
• Crystallography

Background:

• High-resolution biomacromolecular structure determination is crucial for understanding protein function.
• Serial crystallography faces limitations in sample volume and X-ray beamtime access.
• Efficient crystal growth and radiation damage mitigation are key challenges.

Purpose of the Study:

• To introduce an alternative method for biomacromolecular structure determination.
• To present the first ambient temperature lysozyme structure using a home X-ray source.
• To demonstrate the utility of a plate-reader module for rapid data acquisition.

Main Methods:

• Adaptation of a plate-reader module for a 72-well Terasaki plate.
• Utilizing a home X-ray source for structure determination.
• Collecting data at ambient temperature for lysozyme structure determination at the Turkish light source (Turkish DeLight).

Main Results:

• A complete dataset for lysozyme was collected in 18.5 minutes at 2.39 Å resolution.
• The ambient temperature structure provides insights into lysozyme structural dynamics when compared to a cryogenic structure.
• The Turkish DeLight source enabled robust and rapid ambient temperature structure determination with minimal radiation damage.

Conclusions:

• The plate-reader module offers a convenient alternative for biomacromolecular structure determination.
• Ambient temperature data collection provides valuable complementary information to cryogenic data.
• The developed method facilitates efficient structural studies with reduced radiation damage.