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Combination of X-ray crystallography, SAXS and DEER to obtain the structure of the FnIII-3,4 domains of integrin α6β4
Integrin α6β4 is a major component of hemidesmosomes that mediate the stable anchorage of epithelial cells to the underlying basement membrane. Integrin α6β4 has also been implicated in cell proliferation and migration and in carcinoma progression. The third and fourth fibronectin type III domains (...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4388270/ https://www.ncbi.nlm.nih.gov/pubmed/25849406 http://dx.doi.org/10.1107/S1399004715002485 |
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author | Alonso-García, Noelia García-Rubio, Inés Manso, José A. Buey, Rubén M. Urien, Hector Sonnenberg, Arnoud Jeschke, Gunnar de Pereda, José M. |
author_facet | Alonso-García, Noelia García-Rubio, Inés Manso, José A. Buey, Rubén M. Urien, Hector Sonnenberg, Arnoud Jeschke, Gunnar de Pereda, José M. |
author_sort | Alonso-García, Noelia |
collection | PubMed |
description | Integrin α6β4 is a major component of hemidesmosomes that mediate the stable anchorage of epithelial cells to the underlying basement membrane. Integrin α6β4 has also been implicated in cell proliferation and migration and in carcinoma progression. The third and fourth fibronectin type III domains (FnIII-3,4) of integrin β4 mediate binding to the hemidesmosomal proteins BPAG1e and BPAG2, and participate in signalling. Here, it is demonstrated that X-ray crystallography, small-angle X-ray scattering and double electron–electron resonance (DEER) complement each other to solve the structure of the FnIII-3,4 region. The crystal structures of the individual FnIII-3 and FnIII-4 domains were solved and the relative arrangement of the FnIII domains was elucidated by combining DEER with site-directed spin labelling. Multiple structures of the interdomain linker were modelled by Monte Carlo methods complying with DEER constraints, and the final structures were selected against experimental scattering data. FnIII-3,4 has a compact and cambered flat structure with an evolutionary conserved surface that is likely to correspond to a protein-interaction site. Finally, this hybrid method is of general application for the study of other macromolecules and complexes. |
format | Online Article Text |
id | pubmed-4388270 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-43882702015-05-06 Combination of X-ray crystallography, SAXS and DEER to obtain the structure of the FnIII-3,4 domains of integrin α6β4 Alonso-García, Noelia García-Rubio, Inés Manso, José A. Buey, Rubén M. Urien, Hector Sonnenberg, Arnoud Jeschke, Gunnar de Pereda, José M. Acta Crystallogr D Biol Crystallogr Research Papers Integrin α6β4 is a major component of hemidesmosomes that mediate the stable anchorage of epithelial cells to the underlying basement membrane. Integrin α6β4 has also been implicated in cell proliferation and migration and in carcinoma progression. The third and fourth fibronectin type III domains (FnIII-3,4) of integrin β4 mediate binding to the hemidesmosomal proteins BPAG1e and BPAG2, and participate in signalling. Here, it is demonstrated that X-ray crystallography, small-angle X-ray scattering and double electron–electron resonance (DEER) complement each other to solve the structure of the FnIII-3,4 region. The crystal structures of the individual FnIII-3 and FnIII-4 domains were solved and the relative arrangement of the FnIII domains was elucidated by combining DEER with site-directed spin labelling. Multiple structures of the interdomain linker were modelled by Monte Carlo methods complying with DEER constraints, and the final structures were selected against experimental scattering data. FnIII-3,4 has a compact and cambered flat structure with an evolutionary conserved surface that is likely to correspond to a protein-interaction site. Finally, this hybrid method is of general application for the study of other macromolecules and complexes. International Union of Crystallography 2015-03-27 /pmc/articles/PMC4388270/ /pubmed/25849406 http://dx.doi.org/10.1107/S1399004715002485 Text en © Alonso-García et al. 2015 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 Alonso-García, Noelia García-Rubio, Inés Manso, José A. Buey, Rubén M. Urien, Hector Sonnenberg, Arnoud Jeschke, Gunnar de Pereda, José M. Combination of X-ray crystallography, SAXS and DEER to obtain the structure of the FnIII-3,4 domains of integrin α6β4 |
title | Combination of X-ray crystallography, SAXS and DEER to obtain the structure of the FnIII-3,4 domains of integrin α6β4 |
title_full | Combination of X-ray crystallography, SAXS and DEER to obtain the structure of the FnIII-3,4 domains of integrin α6β4 |
title_fullStr | Combination of X-ray crystallography, SAXS and DEER to obtain the structure of the FnIII-3,4 domains of integrin α6β4 |
title_full_unstemmed | Combination of X-ray crystallography, SAXS and DEER to obtain the structure of the FnIII-3,4 domains of integrin α6β4 |
title_short | Combination of X-ray crystallography, SAXS and DEER to obtain the structure of the FnIII-3,4 domains of integrin α6β4 |
title_sort | combination of x-ray crystallography, saxs and deer to obtain the structure of the fniii-3,4 domains of integrin α6β4 |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4388270/ https://www.ncbi.nlm.nih.gov/pubmed/25849406 http://dx.doi.org/10.1107/S1399004715002485 |
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