Author:
Stevenson Hilary P.,Lin Guowu,Barnes Christopher O.,Sutkeviciute Ieva,Krzysiak Troy,Weiss Simon C.,Reynolds Shelley,Wu Ying,Nagarajan Veeranagu,Makhov Alexander M.,Lawrence Robert,Lamm Emily,Clark Lisa,Gardella Timothy J.,Hogue Brenda G.,Ogata Craig M.,Ahn Jinwoo,Gronenborn Angela M.,Conway James F.,Vilardaga Jean-Pierre,Cohen Aina E.,Calero Guillermo
Abstract
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.
Publisher
International Union of Crystallography (IUCr)
Cited by
29 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献