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Ab initio structure determination from prion nanocrystals at atomic resolution by MicroED.

Michael R Sawaya1, Jose Rodriguez1, Duilio Cascio1

  • 1Howard Hughes Medical Institute, University of California, Los Angeles, CA 90024-1570; University of California, Los Angeles-Department of Energy Institute, University of California, Los Angeles, CA 90024-1570; Department of Biological Chemistry, University of California, Los Angeles, CA 90024-1570; Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90024-1570; Molecular Biology Institute, University of California, Los Angeles, CA 90024-1570.

Proceedings of the National Academy of Sciences of the United States of America
|September 21, 2016
PubMed
Summary

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Microcrystal electron diffraction (MicroED) enables high-resolution imaging of tiny biological crystals. Direct phasing methods, similar to X-ray diffraction, are now feasible, overcoming previous obstacles for novel macromolecular structure determination.

Area of Science:

  • Structural biology
  • Biophysics
  • Materials science

Background:

  • Conventional crystallography requires large crystals, limiting complex macromolecule studies.
  • Electron diffraction offers high resolution from nanoscale crystals, but faces challenges like dynamical scattering and phase determination.

Purpose of the Study:

  • To demonstrate the feasibility of ab initio phase determination using MicroED for novel macromolecular structures.
  • To address concerns regarding dynamical scattering and its impact on MicroED phasing.

Main Methods:

  • Microcrystal electron diffraction (MicroED) was applied to determine the structures of four amyloid core variants of the Sup35 prion protein.
  • Direct phasing methods, analogous to those used in X-ray diffraction, were employed.
Keywords:
MicroEDelectron diffractionnanocrystalphasingprion

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Main Results:

  • Accurate phases were obtained for the Sup35 prion protein structures using MicroED, even with nanoscale crystals.
  • The study successfully overcame the challenge of dynamical scattering for ab initio phasing.

Conclusions:

  • Direct phasing methods are viable with MicroED, even in the presence of dynamical scattering.
  • MicroED is a powerful technique for determining the structures of novel, difficult-to-crystallize macromolecules.