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TitleProtein structure determination by electron diffraction using a single three-dimensional nanocrystal.
Journal, issue, pagesActa Crystallogr D Struct Biol, Vol. 73, Issue Pt 9, Page 738-748, Year 2017
Publish dateSep 1, 2017
AuthorsM T B Clabbers / E van Genderen / W Wan / E L Wiegers / T Gruene / J P Abrahams /
PubMed AbstractThree-dimensional nanometre-sized crystals of macromolecules currently resist structure elucidation by single-crystal X-ray crystallography. Here, a single nanocrystal with a diffracting volume of ...Three-dimensional nanometre-sized crystals of macromolecules currently resist structure elucidation by single-crystal X-ray crystallography. Here, a single nanocrystal with a diffracting volume of only 0.14 µm, i.e. no more than 6 × 10 unit cells, provided sufficient information to determine the structure of a rare dimeric polymorph of hen egg-white lysozyme by electron crystallography. This is at least an order of magnitude smaller than was previously possible. The molecular-replacement solution, based on a monomeric polyalanine model, provided sufficient phasing power to show side-chain density, and automated model building was used to reconstruct the side chains. Diffraction data were acquired using the rotation method with parallel beam diffraction on a Titan Krios transmission electron microscope equipped with a novel in-house-designed 1024 × 1024 pixel Timepix hybrid pixel detector for low-dose diffraction data collection. Favourable detector characteristics include the ability to accurately discriminate single high-energy electrons from X-rays and count them, fast readout to finely sample reciprocal space and a high dynamic range. This work, together with other recent milestones, suggests that electron crystallography can provide an attractive alternative in determining biological structures.
External linksActa Crystallogr D Struct Biol / PubMed:28876237 / PubMed Central
MethodsEM (electron crystallography)
Resolution2.11 Å
Structure data

PDB-5o4w:
Protein structure determination by electron diffraction using a single three-dimensional nanocrystal
Method: ELECTRON CRYSTALLOGRAPHY / Resolution: 2.11 Å

PDB-5o4x:
Protein structure determination by electron diffraction using a single three-dimensional nanocrystal
Method: ELECTRON CRYSTALLOGRAPHY / Resolution: 2.11 Å

Source
  • gallus gallus (chicken)
KeywordsHYDROLASE / Lysozyme / Nanocrystal

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