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Title: Combination of X-ray crystallography, SAXS and DEER to obtain the structure of the FnIII-3, 4 domains of integrin α6β4

The structure of the FnIII-3, 4 region of integrin β4 was solved using a hybrid approach that combines crystallographic structures, SAXS, DEER and molecular modelling. The structure helps in understanding how integrin β4 might bind to other hemidesmosomal proteins and mediate signalling. 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 surfacemore » 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.« less
Authors:
 [1] ;  [2] ;  [3] ;  [1] ;  [1] ;  [3] ;  [1] ;  [4] ;  [2] ;  [1]
  1. Consejo Superior de Investigaciones Científicas – University of Salamanca, Campus Unamuno, 37007 Salamanca (Spain)
  2. ETH Zürich, Vladimir-Prelog-Weg 2, CH-8093 Zürich (Switzerland)
  3. (Spain)
  4. Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam (Netherlands)
Publication Date:
OSTI Identifier:
22347703
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography; Journal Volume: 71; Journal Issue: Pt 4; Other Information: PMCID: PMC4388270; PMID: 25849406; PUBLISHER-ID: kw5120; OAI: oai:pubmedcentral.nih.gov:4388270; Copyright (c) Alonso-García et al. 2015; 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.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
Denmark
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CELL PROLIFERATION; COMPACTS; CRYSTAL STRUCTURE; CRYSTALLOGRAPHY; CRYSTALS; ELECTRONS; HYBRIDIZATION; INTERACTIONS; MIGRATION; MONTE CARLO METHOD; RESONANCE; SMALL ANGLE SCATTERING; SPIN; SURFACES