TY - JOUR
T1 - Transmission electron microscopy for the evaluation and optimization of crystal growth
AU - Stevenson, Hilary P.
AU - Lin, Guowu
AU - Barnes, Christopher O.
AU - Sutkeviciute, Ieva
AU - Krzysiak, Troy
AU - Weiss, Simon C.
AU - Reynolds, Shelley
AU - Wu, Ying
AU - Nagarajan, Veeranagu
AU - Makhov, Alexander M.
AU - Lawrence, Robert
AU - Lamm, Emily
AU - Clark, Lisa
AU - Gardella, Timothy J.
AU - Hogue, Brenda
AU - Ogata, Craig M.
AU - Ahn, Jinwoo
AU - Gronenborn, Angela M.
AU - Conway, James F.
AU - Vilardaga, Jean Pierre
AU - Cohen, Aina E.
AU - Calero, Guillermo
N1 - Publisher Copyright:
© 2016 International Union of Crystallography.
PY - 2016
Y1 - 2016
N2 - The crystallization of protein samples remains the most significant challenge in structure determination by X-ray crystallography. Here, the effectiveness of transmission electron microscopy (TEM) analysis to aid in the crystallization of biological macromolecules is demonstrated. It was found that the presence of well ordered lattices with higher order Bragg spots, revealed by Fourier analysis of TEM images, is a good predictor of diffraction-quality crystals. Moreover, the use of TEM allowed (i) comparison of lattice quality among crystals from different conditions in crystallization screens; (ii) the detection of crystal pathologies that could contribute to poor X-ray diffraction, including crystal lattice defects, anisotropic diffraction and crystal contamination by heavy protein aggregates and nanocrystal nuclei; (iii) the qualitative estimation of crystal solvent content to explore the effect of lattice dehydration on diffraction and (iv) the selection of high-quality crystal fragments for microseeding experiments to generate reproducibly larger sized crystals. Applications to X-ray free-electron laser (XFEL) and micro-electron diffraction (microED) experiments are also discussed.
AB - The crystallization of protein samples remains the most significant challenge in structure determination by X-ray crystallography. Here, the effectiveness of transmission electron microscopy (TEM) analysis to aid in the crystallization of biological macromolecules is demonstrated. It was found that the presence of well ordered lattices with higher order Bragg spots, revealed by Fourier analysis of TEM images, is a good predictor of diffraction-quality crystals. Moreover, the use of TEM allowed (i) comparison of lattice quality among crystals from different conditions in crystallization screens; (ii) the detection of crystal pathologies that could contribute to poor X-ray diffraction, including crystal lattice defects, anisotropic diffraction and crystal contamination by heavy protein aggregates and nanocrystal nuclei; (iii) the qualitative estimation of crystal solvent content to explore the effect of lattice dehydration on diffraction and (iv) the selection of high-quality crystal fragments for microseeding experiments to generate reproducibly larger sized crystals. Applications to X-ray free-electron laser (XFEL) and micro-electron diffraction (microED) experiments are also discussed.
KW - Crystal optimization
KW - Crystal optimization
KW - Micro-electron diffraction
KW - Nanocrystallography
KW - Structural biology
KW - Transmission electron microscopy
KW - X-ray free-electron lasers
KW - XFELs
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U2 - 10.1107/S2059798316001546
DO - 10.1107/S2059798316001546
M3 - Article
C2 - 27139624
AN - SCOPUS:84974549964
SN - 0907-4449
VL - 72
SP - 603
EP - 615
JO - Acta Crystallographica Section D: Structural Biology
JF - Acta Crystallographica Section D: Structural Biology
IS - 5
ER -