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Release Factors 2 from Escherichia coli and Thermus thermophilus: structural, spectroscopic and microcalorimetric studies

Prokaryotic class I release factors (RFs) respond to mRNA stop codons and terminate protein synthesis. They interact with the ribosomal decoding site and the peptidyl-transferase centre bridging these 75 Å distant ribosomal centres. For this an elongated RF conformation, with partially unfolded core...

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Autores principales: Zoldák, Gabriel, Redecke, Lars, Svergun, Dmitri I., Konarev, Peter V., Voertler, C. Stefan, Dobbek, Holger, Sedlák, Erik, Sprinzl, Mathias
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1849895/
https://www.ncbi.nlm.nih.gov/pubmed/17272297
http://dx.doi.org/10.1093/nar/gkl696
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author Zoldák, Gabriel
Redecke, Lars
Svergun, Dmitri I.
Konarev, Peter V.
Voertler, C. Stefan
Dobbek, Holger
Sedlák, Erik
Sprinzl, Mathias
author_facet Zoldák, Gabriel
Redecke, Lars
Svergun, Dmitri I.
Konarev, Peter V.
Voertler, C. Stefan
Dobbek, Holger
Sedlák, Erik
Sprinzl, Mathias
author_sort Zoldák, Gabriel
collection PubMed
description Prokaryotic class I release factors (RFs) respond to mRNA stop codons and terminate protein synthesis. They interact with the ribosomal decoding site and the peptidyl-transferase centre bridging these 75 Å distant ribosomal centres. For this an elongated RF conformation, with partially unfolded core domains II·III·IV is required, which contrasts the known compact RF crystal structures. The crystal structure of Thermus thermophilus RF2 was determined and compared with solution structure of T. thermophilus and Escherichia coli RF2 by microcalorimetry, circular dichroism spectroscopy and small angle X-ray scattering. The structure of T. thermophilus RF2 in solution at 20°C is predominantly compact like the crystal structure. Thermodynamic analysis point to an initial melting of domain I, which is independent from the melting of the core. The core domains II·III·IV melt cooperatively at the respective physiological temperatures for T. thermophilus and E. coli. Thermodynamic analyses and the X-ray scattering results for T. thermophilus RF2 in solution suggest that the compact conformation of RF2 resembles a physiological state in absence of the ribosome.
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spelling pubmed-18498952007-04-26 Release Factors 2 from Escherichia coli and Thermus thermophilus: structural, spectroscopic and microcalorimetric studies Zoldák, Gabriel Redecke, Lars Svergun, Dmitri I. Konarev, Peter V. Voertler, C. Stefan Dobbek, Holger Sedlák, Erik Sprinzl, Mathias Nucleic Acids Res Structural Biology Prokaryotic class I release factors (RFs) respond to mRNA stop codons and terminate protein synthesis. They interact with the ribosomal decoding site and the peptidyl-transferase centre bridging these 75 Å distant ribosomal centres. For this an elongated RF conformation, with partially unfolded core domains II·III·IV is required, which contrasts the known compact RF crystal structures. The crystal structure of Thermus thermophilus RF2 was determined and compared with solution structure of T. thermophilus and Escherichia coli RF2 by microcalorimetry, circular dichroism spectroscopy and small angle X-ray scattering. The structure of T. thermophilus RF2 in solution at 20°C is predominantly compact like the crystal structure. Thermodynamic analysis point to an initial melting of domain I, which is independent from the melting of the core. The core domains II·III·IV melt cooperatively at the respective physiological temperatures for T. thermophilus and E. coli. Thermodynamic analyses and the X-ray scattering results for T. thermophilus RF2 in solution suggest that the compact conformation of RF2 resembles a physiological state in absence of the ribosome. Oxford University Press 2007-02 2007-02-01 /pmc/articles/PMC1849895/ /pubmed/17272297 http://dx.doi.org/10.1093/nar/gkl696 Text en © 2007 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
Zoldák, Gabriel
Redecke, Lars
Svergun, Dmitri I.
Konarev, Peter V.
Voertler, C. Stefan
Dobbek, Holger
Sedlák, Erik
Sprinzl, Mathias
Release Factors 2 from Escherichia coli and Thermus thermophilus: structural, spectroscopic and microcalorimetric studies
title Release Factors 2 from Escherichia coli and Thermus thermophilus: structural, spectroscopic and microcalorimetric studies
title_full Release Factors 2 from Escherichia coli and Thermus thermophilus: structural, spectroscopic and microcalorimetric studies
title_fullStr Release Factors 2 from Escherichia coli and Thermus thermophilus: structural, spectroscopic and microcalorimetric studies
title_full_unstemmed Release Factors 2 from Escherichia coli and Thermus thermophilus: structural, spectroscopic and microcalorimetric studies
title_short Release Factors 2 from Escherichia coli and Thermus thermophilus: structural, spectroscopic and microcalorimetric studies
title_sort release factors 2 from escherichia coli and thermus thermophilus: structural, spectroscopic and microcalorimetric studies
topic Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1849895/
https://www.ncbi.nlm.nih.gov/pubmed/17272297
http://dx.doi.org/10.1093/nar/gkl696
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