Ribosome engineering to promote new crystal forms
Crystallographic studies of the ribosome have provided molecular details of protein synthesis. However, the crystallization of functional complexes of ribosomes with GTPase translation factors proved to be elusive for a decade after the first ribosome structures were determined. Analysis of the pack...
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sg-ntu-dr.10356-953142023-02-28T17:00:38Z Ribosome engineering to promote new crystal forms Ramakrishnan, V. Selmer, Maria Gao, Yong-Gui Weixlbaumer, Albert School of Biological Sciences DRNTU::Science::Chemistry::Biochemistry DRNTU::Science::Chemistry::Crystallography Crystallographic studies of the ribosome have provided molecular details of protein synthesis. However, the crystallization of functional complexes of ribosomes with GTPase translation factors proved to be elusive for a decade after the first ribosome structures were determined. Analysis of the packing in different 70S ribosome crystal forms revealed that regardless of the species or space group, a contact between ribosomal protein L9 from the large subunit and 16S rRNA in the shoulder of a neighbouring small subunit in the crystal lattice competes with the binding of GTPase elongation factors to this region of 16S rRNA. To prevent the formation of this preferred crystal contact, a mutant strain of Thermus thermophilus, HB8-MRCMSAW1, in which the ribosomal protein L9 gene has been truncated was constructed by homologous recombination. Mutant 70S ribosomes were used to crystallize and solve the structure of the ribosome with EF-G, GDP and fusidic acid in a previously unobserved crystal form. Subsequent work has shown the usefulness of this strain for crystallization of the ribosome with other GTPase factors. Published version 2013-02-27T02:26:47Z 2019-12-06T19:12:24Z 2013-02-27T02:26:47Z 2019-12-06T19:12:24Z 2012 2012 Journal Article Selmer, M., Gao, Y.-G., Weixlbaumer, A., & Ramakrishnan, V. (2012). Ribosome engineering to promote new crystal forms. Acta Crystallographica Section D Biological Crystallography, 68(5), 578-583. 0907-4449 https://hdl.handle.net/10356/95314 http://hdl.handle.net/10220/9266 10.1107/S0907444912006348 22525755 en Acta crystallographica section D biological crystallography © 2012 International Union of Crystallography. This paper was published in Acta Crystallographica Section D Biological Crystallography and is made available as an electronic reprint (preprint) with permission of International Union of Crystallography. The paper can be found at the following official DOI: [http://dx.doi.org/10.1107/S0907444912006348]. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law. application/pdf |
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DRNTU::Science::Chemistry::Biochemistry DRNTU::Science::Chemistry::Crystallography Ramakrishnan, V. Selmer, Maria Gao, Yong-Gui Weixlbaumer, Albert Ribosome engineering to promote new crystal forms |
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Crystallographic studies of the ribosome have provided molecular details of protein synthesis. However, the crystallization of functional complexes of ribosomes with GTPase translation factors proved to be elusive for a decade after the first ribosome structures were determined. Analysis of the packing in different 70S ribosome crystal forms revealed that regardless of the species or space group, a contact between ribosomal protein L9 from the large subunit and 16S rRNA in the shoulder of a neighbouring small subunit in the crystal lattice competes with the binding of GTPase elongation factors to this region of 16S rRNA. To prevent the formation of this preferred crystal contact, a mutant strain of Thermus thermophilus, HB8-MRCMSAW1, in which the ribosomal protein L9 gene has been truncated was constructed by homologous recombination. Mutant 70S ribosomes were used to crystallize and solve the structure of the ribosome with EF-G, GDP and fusidic acid in a previously unobserved crystal form. Subsequent work has shown the usefulness of this strain for crystallization of the ribosome with other GTPase factors. |
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School of Biological Sciences |
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School of Biological Sciences Ramakrishnan, V. Selmer, Maria Gao, Yong-Gui Weixlbaumer, Albert |
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Article |
author |
Ramakrishnan, V. Selmer, Maria Gao, Yong-Gui Weixlbaumer, Albert |
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Ramakrishnan, V. |
title |
Ribosome engineering to promote new crystal forms |
title_short |
Ribosome engineering to promote new crystal forms |
title_full |
Ribosome engineering to promote new crystal forms |
title_fullStr |
Ribosome engineering to promote new crystal forms |
title_full_unstemmed |
Ribosome engineering to promote new crystal forms |
title_sort |
ribosome engineering to promote new crystal forms |
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2013 |
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https://hdl.handle.net/10356/95314 http://hdl.handle.net/10220/9266 |
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