Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/131053
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Type: | Journal article |
Title: | Derivatization of protein crystals with I3C using random microseed matrix screening |
Author: | Truong, J.Q. Nguyen, S. Bruning, J.B. Shearwin, K.E. |
Citation: | Journal of Visualized Experiments, 2021; 2021(167):1-19 |
Publisher: | Journal of Visualized Experiments |
Issue Date: | 2021 |
ISSN: | 1940-087X 1940-087X |
Statement of Responsibility: | Jia Quyen Truong, Stephanie Nguyen, John B. Bruning, Keith E. Shearwin |
Abstract: | Protein structure elucidation using X-ray crystallography requires both high quality diffracting crystals and computational solution of the diffraction phase problem. Novel structures that lack a suitable homology model are often derivatized with heavy atoms to provide experimental phase information. The presented protocol efficiently generates derivatized protein crystals by combining random microseeding matrix screening with derivatization with a heavy atom molecule I3C (5-amino-2,4,6-triiodoisophthalic acid). By incorporating I3C into the crystal lattice, the diffraction phase problem can be efficiently solved using single wavelength anomalous dispersion (SAD) phasing. The equilateral triangle arrangement of iodine atoms in I3C allows for rapid validation of a correct anomalous substructure. This protocol will be useful to structural biologists who solve macromolecular structures using crystallography-based techniques with interest in experimental phasing. |
Keywords: | Animals Chickens Lithium Triiodobenzoic Acids Muramidase Proteins Imaging, Three-Dimensional Crystallography, X-Ray Diffusion Models, Molecular Data Analysis |
Rights: | © 2021 JoVE Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License |
DOI: | 10.3791/61894 |
Grant ID: | http://purl.org/au-research/grants/arc/DP160101450 http://purl.org/au-research/grants/arc/DP150103009 |
Published version: | http://dx.doi.org/10.3791/61894 |
Appears in Collections: | Aurora harvest 8 Environment Institute publications |
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hdl_131053.pdf | 1.01 MB | Adobe PDF | View/Open |
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