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Viscosity-adjustable grease matrices for serial nanocrystallography.

Michihiro Sugahara1, Koji Motomura2, Mamoru Suzuki3

  • 1RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo, 679-5148, Japan. msuga@spring8.or.jp.

Scientific Reports
|January 30, 2020
PubMed
Summary
This summary is machine-generated.

This study introduces novel viscous grease matrices for serial femtosecond crystallography (SFX), enabling protein structure determination with minimal sample consumption and improved stream stability. These new carriers overcome limitations of conventional greases for X-ray free-electron laser experiments.

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Area of Science:

  • Structural Biology
  • Biophysics
  • Crystallography

Background:

  • Serial femtosecond crystallography (SFX) uses X-ray free-electron lasers to determine protein structures at room temperature with minimal radiation damage.
  • Effective sample delivery is crucial for SFX, requiring stable streams of microcrystals.
  • Conventional grease matrices often exhibit unstable viscosity, leading to stream curling and inconsistent sample loading.

Purpose of the Study:

  • To develop novel viscous grease matrices for improved serial sample loading in SFX.
  • To overcome the limitations of conventional greases, such as unpredictable viscosity and potential protein inhibition.
  • To enable efficient protein structure determination from nano-/microcrystals with reduced sample consumption.

Main Methods:

  • Introduction of two novel grease matrices with known compounds and higher viscosity into the X-ray free-electron laser beam path.
  • Utilizing a high-viscosity micro-extrusion injector for serial sample loading at a low flow rate (~0.5 μl/min).
  • Application of the grease matrices as crystal carriers for serial sample loading.

Main Results:

  • The novel grease matrices provided a continuous and stable sample stream, essential for serial sample loading.
  • Successful determination of the proteinase K structure from nano-/microcrystals was achieved.
  • The method demonstrated the possibility of determining protein structures with less than 1 mg of sample.

Conclusions:

  • Novel viscous grease matrices offer a significant improvement for serial sample loading in SFX.
  • These matrices ensure stable sample streams, reduce sample consumption, and avoid protein inhibition.
  • The developed method advances the capabilities of SFX for structural biology research.