Kilohertz serial crystallography with the JUNGFRAU detector at a fourth-generation synchrotron source

Area of Science:

• Structural Biology
• Biophysics
• Crystallography

Background:

• Serial and time-resolved macromolecular crystallography are advancing rapidly.
• Limited beam time at X-ray free-electron lasers and inadequate infrastructure at synchrotrons pose challenges.
• There is a need for efficient data collection methods at synchrotrons.

Purpose of the Study:

• To demonstrate a new setup for kilohertz serial crystallography data collection at fourth-generation synchrotrons.
• To enable time-resolved protein dynamics studies at microsecond to second timescales.
• To improve accessibility of advanced crystallography techniques for researchers.

Main Methods:

• Utilized the JUNGFRAU detector and Jungfraujoch data-acquisition system.
• Collected kilohertz serial crystallography data at a synchrotron.
• Performed time-resolved experiments on membrane protein KR2 and lysozyme microcrystals.

Main Results:

• Achieved kilohertz data collection rates for serial crystallography.
• Demonstrated the capability to collect multiple-time-point time-resolved protein dynamics data.
• Obtained a complete X-ray dataset from lysozyme microcrystals in 1 minute.
• Showcased microsecond to second dynamics probing at synchrotrons.
• Resolved the dynamics of the sodium-pump KR2 with 1 ms time resolution.

Conclusions:

• The new setup facilitates high-throughput serial and time-resolved crystallography at synchrotrons.
• This technology significantly reduces the time required for probing protein dynamics.
• Future automation of data handling and analysis will enhance accessibility for the scientific community.