5O4X

Protein structure determination by electron diffraction using a single three-dimensional nanocrystal

Summary for 5O4X

• Entry DOI: 10.2210/pdb5o4x/pdb
• Descriptor: Lysozyme C (1 entity in total)
• Functional Keywords: lysozyme, nanocrystal, hydrolase
• Biological source: Gallus gallus (Chicken)
• Cellular location: Secreted: P00698
• Total number of polymer chains: 2
• Total formula weight: 28662.32

Authors

Clabbers, M.T.B.,van Genderen, E.,Wan, W.,Wiegers, E.L.,Gruene, T.,Abrahams, J.P. (deposition date: 2017-05-31, release date: 2017-08-23, Last modification date: 2024-01-17)

Primary citation

Clabbers, M.T.B.,van Genderen, E.,Wan, W.,Wiegers, E.L.,Gruene, T.,Abrahams, J.P.
Protein structure determination by electron diffraction using a single three-dimensional nanocrystal.
Acta Crystallogr D Struct Biol, 73:738-748, 2017

PubMed Abstract: 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.

PubMed: 28876237
DOI: 10.1107/S2059798317010348

Experimental method

ELECTRON CRYSTALLOGRAPHY (2.11 Å)