Fixed-target serial crystallography at the Structural Biology Center.

Area of Science:

• Structural biology
• Biophysics
• Crystallography

Background:

• Serial synchrotron crystallography (SSC) is crucial for studying protein structures under physiological conditions.
• Existing SSC methods face challenges with sample handling and radiation damage.
• Advancements are needed for efficient data collection from numerous crystals.

Purpose of the Study:

• To introduce a novel 3D-printed mesh-holder for SSC.
• To optimize sample immobilization and handling for X-ray crystallography.
• To apply the system to study important beta-lactamase enzymes.

Main Methods:

• Development of a 3D-printed mesh-holder for immobilizing crystal suspensions.
• Utilizing a fixed-target approach with Mylar films for crystal trapping.
• Employing piezoelectric stages and a high-performance computing pipeline for data acquisition and analysis.

Main Results:

• The new holder effectively immobilizes near-monolayers of crystals in their mother liquor.
• Rapid data collection in scan mode was achieved.
• The system was successfully applied to determine structures of two beta-lactamases.

Conclusions:

• The 3D-printed mesh-holder system enhances serial synchrotron crystallography efficiency.
• This method facilitates near real-time analysis of protein structures.
• The system is applicable to diverse protein targets, including clinically relevant enzymes.