2024-03-29T09:32:07Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/1348322021-12-28T16:27:30Zcom_10261_112com_10261_1col_10261_365
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.
Martínez Buey, Rubén
Urien, Hector
Sonnenberg, Arnoud
Jeschke, Gunnar
Pereda, José M. de
Consejo Superior de Investigaciones Científicas (España)
Swiss National Science Foundation
Dutch Cancer Society
European Commission
Ministerio de Economía y Competitividad (España)
Netherlands Organization for Scientific Research
Universidad de Salamanca
Integrin 64
Double electron–electron resonance
FnIII-4
Smallangle X-ray scattering
FnIII-3
This is an open-access article distributed under the terms of the Creative Commons Attribution Licence.
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.
2016-07-15T11:13:38Z
2016-07-15T11:13:38Z
2015
2016-07-15T11:13:40Z
artículo
Acta Crystallographica Section D: Biological Crystallography 71: 969-985 (2015)
http://hdl.handle.net/10261/134832
10.1107/S1399004715002485
http://dx.doi.org/10.13039/501100003339
http://dx.doi.org/10.13039/501100000780
http://dx.doi.org/10.13039/501100003329
25849406
eng
Publisher's version
http://dx.doi.org/10.1107/S1399004715002485
Sí
info:eu-repo/grantAgreement/EC/FP7/283570
openAccess
International Union of Crystallography