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Volume 26, Issue 4, Pages e3 (April 2018)

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1 Volume 26, Issue 4, Pages 667-675.e3 (April 2018)
A Rare Lysozyme Crystal Form Solved Using Highly Redundant Multiple Electron Diffraction Datasets from Micron-Sized Crystals  Hongyi Xu, Hugo Lebrette, Taimin Yang, Vivek Srinivas, Sven Hovmöller, Martin Högbom, Xiaodong Zou  Structure  Volume 26, Issue 4, Pages e3 (April 2018) DOI: /j.str Copyright © 2018 Elsevier Ltd Terms and Conditions

2 Structure 2018 26, 667-675.e3DOI: (10.1016/j.str.2018.02.015)
Copyright © 2018 Elsevier Ltd Terms and Conditions

3 Figure 1 Lysozyme Crystals and ED Pattern
(A) An optical image of the as-grown crystals. Scale bar, 100 μm. (B) A low-magnification TEM image showing vitrified needle-shaped lysozyme crystals on a TEM grid. Scale bar, 2 μm. (C) A typical ED frame taken from a needle-shaped lysozyme crystal. A diffraction spot with a d spacing of 2 Å is indicated. Structure  , e3DOI: ( /j.str ) Copyright © 2018 Elsevier Ltd Terms and Conditions

4 Figure 2 Data Completeness and Omit Maps
Top: reflections in reciprocal space showing the data completeness from (A) 1-crystal dataset, (B) 4-crystal dataset, and (C) 33-crystal dataset. Reflections are displayed as spheres and colored in a rainbow gradient. Missing reflections are in white and systematic absences in pink (prepared with Reflection Data Viewer in Phenix). Bottom: simulated annealing omit Fo − Fc electrostatic potential map (contoured to 2.8σ, colored in green) calculated after deletion of an entire α helix from residue 26 to 35 in both chains of the asymmetric unit from (A) 1-crystal dataset, (B) 4-crystal dataset, and (C) 33-crystal dataset. Carbon, oxygen, and nitrogen atoms are colored gray, red, and blue, respectively. Structure  , e3DOI: ( /j.str ) Copyright © 2018 Elsevier Ltd Terms and Conditions

5 Figure 3 Improvement of Detail Definition in the Electrostatic Potential Map Simulated annealing 2Fo − Fc and omit Fo − Fc electrostatic potential maps (contoured to 1σ and 3σ and colored in blue and green, respectively), calculated after deletion of water molecules and ions and superimposed on the final models refined against 1-crystal, 4-crystal, 10-best crystal, 10-random crystal, 20-best crystal, and 33-crystal datasets, respectively. Left columns: two water molecules and a probable alternative conformation for a glutamine can be observed only in the map obtained from the 33-crystal dataset. Right columns: a Na+ ion can be modeled only in the map obtained from the 33-crystal dataset, suggested by a higher peak in the omit Fo − Fc map and the presence of two additional water molecules completing its octahedral geometry coordination. Carbon, oxygen, and nitrogen atoms are colored gray, red, and blue, respectively; water molecules and the Na+ ion are represented as red and purple spheres, respectively. Structure  , e3DOI: ( /j.str ) Copyright © 2018 Elsevier Ltd Terms and Conditions


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