Serial protein crystallography in an electron microscope

Nat Commun. 2020 Feb 21;11(1):996. doi: 10.1038/s41467-020-14793-0.

Abstract

Serial X-ray crystallography at free-electron lasers allows to solve biomolecular structures from sub-micron-sized crystals. However, beam time at these facilities is scarce, and involved sample delivery techniques are required. On the other hand, rotation electron diffraction (MicroED) has shown great potential as an alternative means for protein nano-crystallography. Here, we present a method for serial electron diffraction of protein nanocrystals combining the benefits of both approaches. In a scanning transmission electron microscope, crystals randomly dispersed on a sample grid are automatically mapped, and a diffraction pattern at fixed orientation is recorded from each at a high acquisition rate. Dose fractionation ensures minimal radiation damage effects. We demonstrate the method by solving the structure of granulovirus occlusion bodies and lysozyme to resolutions of 1.55 Å and 1.80 Å, respectively. Our method promises to provide rapid structure determination for many classes of materials with minimal sample consumption, using readily available instrumentation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallography / methods*
  • Microscopy, Electron, Scanning Transmission
  • Models, Molecular
  • Muramidase / chemistry
  • Muramidase / ultrastructure
  • Nanoparticles / chemistry
  • Nanoparticles / ultrastructure
  • Occlusion Body Matrix Proteins / chemistry
  • Occlusion Body Matrix Proteins / ultrastructure
  • Particle Size
  • Protein Conformation
  • Proteins / chemistry*
  • Proteins / ultrastructure

Substances

  • Occlusion Body Matrix Proteins
  • Proteins
  • granulin protein, granulovirus
  • Muramidase