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A description of the structural determination procedures of a gap junction channel at 3.5 Å resolution

Intercellular signalling is an essential characteristic of multicellular organisms. Gap junctions, which consist of arrays of intercellular channels, permit the exchange of ions and small molecules between adjacent cells. Here, the structural deter­mination of a gap junction channel composed of conn...

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Autores principales: Suga, Michihiro, Maeda, Shoji, Nakagawa, So, Yamashita, Eiki, Tsukihara, Tomitake
Formato: Texto
Lenguaje:English
Publicado: International Union of Crystallography 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2714718/
https://www.ncbi.nlm.nih.gov/pubmed/19622859
http://dx.doi.org/10.1107/S0907444909014711
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author Suga, Michihiro
Maeda, Shoji
Nakagawa, So
Yamashita, Eiki
Tsukihara, Tomitake
author_facet Suga, Michihiro
Maeda, Shoji
Nakagawa, So
Yamashita, Eiki
Tsukihara, Tomitake
author_sort Suga, Michihiro
collection PubMed
description Intercellular signalling is an essential characteristic of multicellular organisms. Gap junctions, which consist of arrays of intercellular channels, permit the exchange of ions and small molecules between adjacent cells. Here, the structural deter­mination of a gap junction channel composed of connexin 26 (Cx26) at 3.5 Å resolution is described. During each step of the purification process, the protein was examined using electron microscopy and/or dynamic light scattering. Dehydration of the crystals improved the resolution limits. Phase refinement using multi-crystal averaging in conjunction with noncrystallographic symmetry averaging based on strictly determined noncrystallographic symmetry operators resulted in an electron-density map for model building. The amino-acid sequence of a protomer structure consisting of the amino-terminal helix, four transmembrane helices and two extracellular loops was assigned to the electron-density map. The amino-acid assignment was confirmed using six selenomethionine (SeMet) sites in the difference Fourier map of the SeMet derivative and three intramolecular disulfide bonds in the anomalous difference Fourier map of the native crystal.
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spelling pubmed-27147182009-07-24 A description of the structural determination procedures of a gap junction channel at 3.5 Å resolution Suga, Michihiro Maeda, Shoji Nakagawa, So Yamashita, Eiki Tsukihara, Tomitake Acta Crystallogr D Biol Crystallogr Research Papers Intercellular signalling is an essential characteristic of multicellular organisms. Gap junctions, which consist of arrays of intercellular channels, permit the exchange of ions and small molecules between adjacent cells. Here, the structural deter­mination of a gap junction channel composed of connexin 26 (Cx26) at 3.5 Å resolution is described. During each step of the purification process, the protein was examined using electron microscopy and/or dynamic light scattering. Dehydration of the crystals improved the resolution limits. Phase refinement using multi-crystal averaging in conjunction with noncrystallographic symmetry averaging based on strictly determined noncrystallographic symmetry operators resulted in an electron-density map for model building. The amino-acid sequence of a protomer structure consisting of the amino-terminal helix, four transmembrane helices and two extracellular loops was assigned to the electron-density map. The amino-acid assignment was confirmed using six selenomethionine (SeMet) sites in the difference Fourier map of the SeMet derivative and three intramolecular disulfide bonds in the anomalous difference Fourier map of the native crystal. International Union of Crystallography 2009-08-01 2009-07-10 /pmc/articles/PMC2714718/ /pubmed/19622859 http://dx.doi.org/10.1107/S0907444909014711 Text en © Suga et al. 2009 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.
spellingShingle Research Papers
Suga, Michihiro
Maeda, Shoji
Nakagawa, So
Yamashita, Eiki
Tsukihara, Tomitake
A description of the structural determination procedures of a gap junction channel at 3.5 Å resolution
title A description of the structural determination procedures of a gap junction channel at 3.5 Å resolution
title_full A description of the structural determination procedures of a gap junction channel at 3.5 Å resolution
title_fullStr A description of the structural determination procedures of a gap junction channel at 3.5 Å resolution
title_full_unstemmed A description of the structural determination procedures of a gap junction channel at 3.5 Å resolution
title_short A description of the structural determination procedures of a gap junction channel at 3.5 Å resolution
title_sort description of the structural determination procedures of a gap junction channel at 3.5 å resolution
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2714718/
https://www.ncbi.nlm.nih.gov/pubmed/19622859
http://dx.doi.org/10.1107/S0907444909014711
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