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Selenium derivatization of nucleic acids for crystallography
The high-resolution structure of the DNA (5′-GTGTACA-C-3′) with the selenium derivatization at the 2′-position of T2 was determined via MAD and SAD phasing. The selenium-derivatized structure (1.28 Å resolution) with the 2′-Se modification in the minor groove is isomorphorous to the native structure...
Autores principales: | , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2007
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1802610/ https://www.ncbi.nlm.nih.gov/pubmed/17169989 http://dx.doi.org/10.1093/nar/gkl1070 |
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author | Jiang, Jiansheng Sheng, Jia Carrasco, Nicolas Huang, Zhen |
author_facet | Jiang, Jiansheng Sheng, Jia Carrasco, Nicolas Huang, Zhen |
author_sort | Jiang, Jiansheng |
collection | PubMed |
description | The high-resolution structure of the DNA (5′-GTGTACA-C-3′) with the selenium derivatization at the 2′-position of T2 was determined via MAD and SAD phasing. The selenium-derivatized structure (1.28 Å resolution) with the 2′-Se modification in the minor groove is isomorphorous to the native structure (2.0 Å). To directly compare with the conventional bromine derivatization, we incorporated bromine into the 5-postion of T4, determined the bromine-derivatized DNA structure at 1.5 Å resolution, and found that the local backbone torsion angles and solvent hydration patterns were altered in the structure with the Br incorporation in the major groove. Furthermore, while the native and Br-derivatized DNAs needed over a week to form reasonable-size crystals, we observed that the Se-derivatized DNAs grew crystals overnight with high-diffraction quality, suggesting that the Se derivatization facilitated the crystal formation. In addition, the Se-derivatized DNA sequences crystallized under a broader range of buffer conditions, and generally had a faster crystal growth rate. Our experimental results indicate that the selenium derivatization of DNAs may facilitate the determination of nucleic acid X-ray crystal structures in phasing and high-quality crystal growth. In addition, our results suggest that the Se derivatization can be an alternative to the conventional Br derivatization. |
format | Text |
id | pubmed-1802610 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2007 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-18026102007-03-01 Selenium derivatization of nucleic acids for crystallography Jiang, Jiansheng Sheng, Jia Carrasco, Nicolas Huang, Zhen Nucleic Acids Res Structural Biology The high-resolution structure of the DNA (5′-GTGTACA-C-3′) with the selenium derivatization at the 2′-position of T2 was determined via MAD and SAD phasing. The selenium-derivatized structure (1.28 Å resolution) with the 2′-Se modification in the minor groove is isomorphorous to the native structure (2.0 Å). To directly compare with the conventional bromine derivatization, we incorporated bromine into the 5-postion of T4, determined the bromine-derivatized DNA structure at 1.5 Å resolution, and found that the local backbone torsion angles and solvent hydration patterns were altered in the structure with the Br incorporation in the major groove. Furthermore, while the native and Br-derivatized DNAs needed over a week to form reasonable-size crystals, we observed that the Se-derivatized DNAs grew crystals overnight with high-diffraction quality, suggesting that the Se derivatization facilitated the crystal formation. In addition, the Se-derivatized DNA sequences crystallized under a broader range of buffer conditions, and generally had a faster crystal growth rate. Our experimental results indicate that the selenium derivatization of DNAs may facilitate the determination of nucleic acid X-ray crystal structures in phasing and high-quality crystal growth. In addition, our results suggest that the Se derivatization can be an alternative to the conventional Br derivatization. Oxford University Press 2007-01 2006-12-14 /pmc/articles/PMC1802610/ /pubmed/17169989 http://dx.doi.org/10.1093/nar/gkl1070 Text en © 2006 The Author(s) This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Structural Biology Jiang, Jiansheng Sheng, Jia Carrasco, Nicolas Huang, Zhen Selenium derivatization of nucleic acids for crystallography |
title | Selenium derivatization of nucleic acids for crystallography |
title_full | Selenium derivatization of nucleic acids for crystallography |
title_fullStr | Selenium derivatization of nucleic acids for crystallography |
title_full_unstemmed | Selenium derivatization of nucleic acids for crystallography |
title_short | Selenium derivatization of nucleic acids for crystallography |
title_sort | selenium derivatization of nucleic acids for crystallography |
topic | Structural Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1802610/ https://www.ncbi.nlm.nih.gov/pubmed/17169989 http://dx.doi.org/10.1093/nar/gkl1070 |
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