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Deriving the ultrastructure of α-crustacyanin using lower-resolution structural and biophysical methods

The low-resolution structure of α-crustacyanin has been determined to 30 Å resolution using negative-stain electron microscopy (EM) with single-particle averaging. The protein, which is an assembly of eight β-crustacyanin dimers, appears asymmetrical and rather open in layout. A model was built to t...

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Autores principales: Rhys, Natasha H., Wang, Ming-Chuan, Jowitt, Thomas A., Helliwell, John R., Grossmann, J. Günter, Baldock, Clair
Formato: Texto
Lenguaje:English
Publicado: International Union of Crystallography 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3004261/
https://www.ncbi.nlm.nih.gov/pubmed/21169698
http://dx.doi.org/10.1107/S0909049510034977
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author Rhys, Natasha H.
Wang, Ming-Chuan
Jowitt, Thomas A.
Helliwell, John R.
Grossmann, J. Günter
Baldock, Clair
author_facet Rhys, Natasha H.
Wang, Ming-Chuan
Jowitt, Thomas A.
Helliwell, John R.
Grossmann, J. Günter
Baldock, Clair
author_sort Rhys, Natasha H.
collection PubMed
description The low-resolution structure of α-crustacyanin has been determined to 30 Å resolution using negative-stain electron microscopy (EM) with single-particle averaging. The protein, which is an assembly of eight β-crustacyanin dimers, appears asymmetrical and rather open in layout. A model was built to the EM map using the X-ray crystallographic structure of β-crustacyanin guided by PISA interface analyses. The model has a theoretical sedimentation coefficient that matches well with the experimentally derived value from sedimentation velocity analytical ultracentrifugation. Additionally, the EM model has similarities to models calculated independently by rigid-body modelling to small-angle X-ray scattering (SAXS) data and extracted in silico from the β-crustacyanin crystal lattice. Theoretical X-ray scattering from each of these models is in reasonable agreement with the experimental SAXS data and together suggest an overall design for the α-crustacyanin assembly.
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spelling pubmed-30042612010-12-23 Deriving the ultrastructure of α-crustacyanin using lower-resolution structural and biophysical methods Rhys, Natasha H. Wang, Ming-Chuan Jowitt, Thomas A. Helliwell, John R. Grossmann, J. Günter Baldock, Clair J Synchrotron Radiat Diffraction Structural Biology The low-resolution structure of α-crustacyanin has been determined to 30 Å resolution using negative-stain electron microscopy (EM) with single-particle averaging. The protein, which is an assembly of eight β-crustacyanin dimers, appears asymmetrical and rather open in layout. A model was built to the EM map using the X-ray crystallographic structure of β-crustacyanin guided by PISA interface analyses. The model has a theoretical sedimentation coefficient that matches well with the experimentally derived value from sedimentation velocity analytical ultracentrifugation. Additionally, the EM model has similarities to models calculated independently by rigid-body modelling to small-angle X-ray scattering (SAXS) data and extracted in silico from the β-crustacyanin crystal lattice. Theoretical X-ray scattering from each of these models is in reasonable agreement with the experimental SAXS data and together suggest an overall design for the α-crustacyanin assembly. International Union of Crystallography 2011-01-01 2010-11-05 /pmc/articles/PMC3004261/ /pubmed/21169698 http://dx.doi.org/10.1107/S0909049510034977 Text en © Natasha H. Rhys et al. 2011 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 Diffraction Structural Biology
Rhys, Natasha H.
Wang, Ming-Chuan
Jowitt, Thomas A.
Helliwell, John R.
Grossmann, J. Günter
Baldock, Clair
Deriving the ultrastructure of α-crustacyanin using lower-resolution structural and biophysical methods
title Deriving the ultrastructure of α-crustacyanin using lower-resolution structural and biophysical methods
title_full Deriving the ultrastructure of α-crustacyanin using lower-resolution structural and biophysical methods
title_fullStr Deriving the ultrastructure of α-crustacyanin using lower-resolution structural and biophysical methods
title_full_unstemmed Deriving the ultrastructure of α-crustacyanin using lower-resolution structural and biophysical methods
title_short Deriving the ultrastructure of α-crustacyanin using lower-resolution structural and biophysical methods
title_sort deriving the ultrastructure of α-crustacyanin using lower-resolution structural and biophysical methods
topic Diffraction Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3004261/
https://www.ncbi.nlm.nih.gov/pubmed/21169698
http://dx.doi.org/10.1107/S0909049510034977
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