A data set from flash X-ray imaging of carboxysomes

Area of Science:

• Structural biology
• Biophysics
• Biochemistry

Background:

• Carboxysomes are essential organelles for carbon fixation in cyanobacteria and proteobacteria.
• Their inherent size heterogeneity and polyhedral structure make traditional crystallization methods challenging.
• Understanding carboxysome structure is key to comprehending their role in Earth's carbon cycle.

Purpose of the Study:

• To investigate the structural heterogeneity of carboxysome particles using advanced X-ray techniques.
• To demonstrate the capability of femtosecond X-ray pulses for analyzing non-crystalline biological samples.
• To develop high-throughput methods for studying structural variations in biological macromolecules.

Main Methods:

• Utilized ultra-intense femtosecond X-ray pulses from X-ray lasers for diffraction studies.
• Employed an aerosol sample injector for high-throughput data collection.
• Recorded 70,000 low-noise diffraction patterns from carboxysome particles within 12 minutes.
• Applied direct phasing methods to separate and analyze individual diffraction patterns.

Main Results:

• Successfully obtained a large dataset of diffraction patterns from heterogeneous carboxysome particles.
• Demonstrated the ability to resolve structural variations within the carboxysome population.
• Characterized carboxysomes with a mean diameter of 115±26 nm from Halothiobacillus neapolitanus.

Conclusions:

• Femtosecond X-ray diffraction offers a powerful, high-throughput approach for studying the structure of single, non-crystalline biological particles.
• This method effectively captures and analyzes structural heterogeneity, opening new avenues in structural biology.
• The findings pave the way for accurate structural studies on a wide range of biological systems previously inaccessible to crystallography.