A Pseudo-Atomic Model of the COPII Cage Obtained from CryoEM and Mass Spectrometry Analyses

COPII vesicles transport proteins from the ER to the Golgi apparatus. Previous cryoEM structures of the COPII cage lacked the resolution necessary to determine the residues of Sec13 and Sec31 that mediate assembly and flexibility of the COPII cage. Here we present a 12Å-resolution structure of the C...

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Autores principales: Noble, Alex J., Zhang, Qian, O’Donnell, Jason, Hariri, Hanaa, Bhattacharya, Nilakshee, Marshall, Alan G., Stagg, Scott M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2012
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3565055/
https://www.ncbi.nlm.nih.gov/pubmed/23262493
http://dx.doi.org/10.1038/nsmb.2467
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author Noble, Alex J.
Zhang, Qian
O’Donnell, Jason
Hariri, Hanaa
Bhattacharya, Nilakshee
Marshall, Alan G.
Stagg, Scott M.
author_facet Noble, Alex J.
Zhang, Qian
O’Donnell, Jason
Hariri, Hanaa
Bhattacharya, Nilakshee
Marshall, Alan G.
Stagg, Scott M.
author_sort Noble, Alex J.
collection PubMed
description COPII vesicles transport proteins from the ER to the Golgi apparatus. Previous cryoEM structures of the COPII cage lacked the resolution necessary to determine the residues of Sec13 and Sec31 that mediate assembly and flexibility of the COPII cage. Here we present a 12Å-resolution structure of the COPII cage, where the tertiary structure of Sec13 and Sec31 is clearly identifiable. We employ this structure and a homology model of the Sec13-Sec31 complex to create a reliable pseudo-atomic model of the COPII cage. We combined this model with hydrogen/deuterium exchange mass spectrometry analysis to characterize four distinct contact regions at the vertices of the COPII cage. Furthermore, we found that the 2-fold symmetry of the Sec31 dimeric region of Sec13-31 is broken on cage formation, and that the resulting hinge is essential to form the proper edge geometry in COPII cages.
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spelling pubmed-35650552013-08-01 A Pseudo-Atomic Model of the COPII Cage Obtained from CryoEM and Mass Spectrometry Analyses Noble, Alex J. Zhang, Qian O’Donnell, Jason Hariri, Hanaa Bhattacharya, Nilakshee Marshall, Alan G. Stagg, Scott M. Nat Struct Mol Biol Article COPII vesicles transport proteins from the ER to the Golgi apparatus. Previous cryoEM structures of the COPII cage lacked the resolution necessary to determine the residues of Sec13 and Sec31 that mediate assembly and flexibility of the COPII cage. Here we present a 12Å-resolution structure of the COPII cage, where the tertiary structure of Sec13 and Sec31 is clearly identifiable. We employ this structure and a homology model of the Sec13-Sec31 complex to create a reliable pseudo-atomic model of the COPII cage. We combined this model with hydrogen/deuterium exchange mass spectrometry analysis to characterize four distinct contact regions at the vertices of the COPII cage. Furthermore, we found that the 2-fold symmetry of the Sec31 dimeric region of Sec13-31 is broken on cage formation, and that the resulting hinge is essential to form the proper edge geometry in COPII cages. 2012-12-23 2013-02 /pmc/articles/PMC3565055/ /pubmed/23262493 http://dx.doi.org/10.1038/nsmb.2467 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Noble, Alex J.
Zhang, Qian
O’Donnell, Jason
Hariri, Hanaa
Bhattacharya, Nilakshee
Marshall, Alan G.
Stagg, Scott M.
A Pseudo-Atomic Model of the COPII Cage Obtained from CryoEM and Mass Spectrometry Analyses
title A Pseudo-Atomic Model of the COPII Cage Obtained from CryoEM and Mass Spectrometry Analyses
title_full A Pseudo-Atomic Model of the COPII Cage Obtained from CryoEM and Mass Spectrometry Analyses
title_fullStr A Pseudo-Atomic Model of the COPII Cage Obtained from CryoEM and Mass Spectrometry Analyses
title_full_unstemmed A Pseudo-Atomic Model of the COPII Cage Obtained from CryoEM and Mass Spectrometry Analyses
title_short A Pseudo-Atomic Model of the COPII Cage Obtained from CryoEM and Mass Spectrometry Analyses
title_sort pseudo-atomic model of the copii cage obtained from cryoem and mass spectrometry analyses
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3565055/
https://www.ncbi.nlm.nih.gov/pubmed/23262493
http://dx.doi.org/10.1038/nsmb.2467
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