Machining protein microcrystals for structure determination by electron diffraction

Proc Natl Acad Sci U S A. 2018 Sep 18;115(38):9569-9573. doi: 10.1073/pnas.1809978115. Epub 2018 Aug 31.

Abstract

We demonstrate that ion-beam milling of frozen, hydrated protein crystals to thin lamella preserves the crystal lattice to near-atomic resolution. This provides a vehicle for protein structure determination, bridging the crystal size gap between the nanometer scale of conventional electron diffraction and micron scale of synchrotron microfocus beamlines. The demonstration that atomic information can be retained suggests that milling could provide such detail on sections cut from vitrified cells.

Keywords: FIB; diffraction; electron crystallography; protein crystal; structure.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cryoelectron Microscopy / methods
  • Crystallography, X-Ray / instrumentation
  • Crystallography, X-Ray / methods*
  • Electrons
  • Ions
  • Microtechnology / instrumentation
  • Microtechnology / methods*
  • Muramidase / chemistry
  • Muramidase / ultrastructure*
  • Synchrotrons

Substances

  • Ions
  • Muramidase

Associated data

  • PDB/6H3B