4.8 Article

Machining protein microcrystals for structure determination by electron diffraction

Publisher

NATL ACAD SCIENCES
DOI: 10.1073/pnas.1809978115

Keywords

electron crystallography; FIB; protein crystal; diffraction; structure

Funding

  1. Wellcome [ALR00750-B500.1, ALR00040, 101122/Z/13/Z, 206422/Z/17/Z, 090532/Z/09/Z]
  2. UK Medical Research Council (MRC) [MR/N00065X/1]
  3. NIH [P41GM103832]
  4. Wellcome
  5. MRC
  6. Biotechnology and Biological Sciences Research Council
  7. BBSRC [BB/S003339/1] Funding Source: UKRI
  8. MRC [MR/N00065X/1] Funding Source: UKRI

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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.

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