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Towards time-resolved serial crystallography in a microfluidic device.

Ashtamurthy S Pawate1, Vukica Šrajer2, Jeremy Schieferstein1

  • 1Department of Chemical and Biomolecular Engineering, The University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

Acta Crystallographica. Section F, Structural Biology Communications
|July 7, 2015
PubMed
Summary
This summary is machine-generated.

Serial crystallography using a microfluidic platform enables dynamic structural studies of proteins. This method captured key states of photoactive yellow protein (PYP) at microsecond and millisecond timescales.

Keywords:
Laue diffractionmicrofluidicsphotoactive yellow proteinprotein crystallizationserial crystallographytime-resolved protein crystallography

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Area of Science:

  • Structural Biology
  • Biophysics
  • Crystallography

Background:

  • Dynamic structural studies are crucial for understanding protein function.
  • Traditional crystallography faces limitations with radiation damage and reaction initiation.
  • Serial crystallography offers a promising alternative for time-resolved studies.

Purpose of the Study:

  • To develop and apply serial time-resolved Laue diffraction for dynamic structural analysis.
  • To investigate the structural intermediates of photoactive yellow protein (PYP).
  • To demonstrate the utility of a microfluidic platform for crystallographic studies.

Main Methods:

  • Utilized a microfluidic crystallization platform for sample handling.
  • Employed serial time-resolved Laue diffraction for data collection.
  • Initiated protein reactions using pulsed laser illumination.

Main Results:

  • Successfully obtained electron-density difference maps.
  • Depicted key structural states (pR(1)/pR(E46Q), pR(2)/pR(CW)) at 10 µs.
  • Identified the pB1 intermediate at 1 ms.

Conclusions:

  • Serial crystallography is effective for studying dynamic protein structures.
  • The microfluidic platform facilitates time-resolved crystallographic analysis.
  • This approach has potential for chemical triggering and multivariable dynamic studies.