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Lipidico Injection Protocol for Serial Crystallography Measurements at the Australian Synchrotron
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The hit-and-return system enables efficient time-resolved serial synchrotron crystallography.

Eike C Schulz1, Pedram Mehrabi1, Henrike M Müller-Werkmeister1,2

  • 1Department for Atomically Resolved Dynamics, Max Planck Institute for Structure and Dynamics of Matter, Hamburg, Germany.

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Area of Science:

  • Structural Biology
  • Biochemistry
  • Crystallography

Background:

  • Time-resolved crystallography is crucial for understanding dynamic biological processes.
  • Capturing transient intermediate structures requires high temporal resolution.
  • Existing methods may have limitations in achieving millisecond to second time scales.

Purpose of the Study:

  • To introduce a novel 'hit-and-return' (HARE) method for time-resolved serial synchrotron crystallography.
  • To enable data collection at millisecond to second time resolutions.
  • To investigate intermediate structures during ligand binding events.

Main Methods:

  • Development of the 'hit-and-return' (HARE) method.
  • Mechanical timing delay system utilizing fixed-target crystallography chips and a translation stage.
  • Optical pump-probe experiments for time-resolved data acquisition.

Main Results:

  • Demonstrated feasibility of HARE for time-resolved serial synchrotron crystallography.
  • Successful data collection at various time intervals: 30 ms, 752 ms, and 2,052 ms.
  • Captured intermediate structures of fluoroacetate dehalogenase during ligand binding.

Conclusions:

  • The HARE method provides a robust approach for time-resolved crystallography.
  • Enables the study of molecular dynamics across a wide range of timescales (milliseconds to seconds).
  • Facilitates the elucidation of reaction mechanisms and intermediate states in biological systems.