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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Main Authors: Ramakrishnan, V., Selmer, Maria, Gao, Yong-Gui, Weixlbaumer, Albert
Other Authors: School of Biological Sciences
Format: Article
Language:English
Published: 2013
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Online Access:https://hdl.handle.net/10356/95314
http://hdl.handle.net/10220/9266
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spelling 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
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Chemistry::Biochemistry
DRNTU::Science::Chemistry::Crystallography
spellingShingle DRNTU::Science::Chemistry::Biochemistry
DRNTU::Science::Chemistry::Crystallography
Ramakrishnan, V.
Selmer, Maria
Gao, Yong-Gui
Weixlbaumer, Albert
Ribosome engineering to promote new crystal forms
description 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.
author2 School of Biological Sciences
author_facet School of Biological Sciences
Ramakrishnan, V.
Selmer, Maria
Gao, Yong-Gui
Weixlbaumer, Albert
format Article
author Ramakrishnan, V.
Selmer, Maria
Gao, Yong-Gui
Weixlbaumer, Albert
author_sort 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
publishDate 2013
url https://hdl.handle.net/10356/95314
http://hdl.handle.net/10220/9266
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