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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 (...

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Autores principales: Alonso-García, Noelia, García-Rubio, Inés, Manso, José A., Buey, Rubén M., Urien, Hector, Sonnenberg, Arnoud, Jeschke, Gunnar, de Pereda, José M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2015
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.
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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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