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TitleCombination of X-ray crystallography, SAXS and DEER to obtain the structure of the FnIII-3,4 domains of integrin α6β4.
Journal, issue, pagesActa Crystallogr D Biol Crystallogr, Vol. 71, Issue Pt 4, Page 969-985, Year 2015
Publish dateMar 27, 2015
AuthorsNoelia Alonso-García / Inés García-Rubio / José A Manso / Rubén M Buey / Hector Urien / Arnoud Sonnenberg / Gunnar Jeschke / José M de Pereda /
PubMed AbstractIntegrin 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 ...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.
External linksActa Crystallogr D Biol Crystallogr / PubMed:25849406 / PubMed Central
MethodsSAS (X-ray synchrotron) / X-ray diffraction
Resolution1.5 - 1.606 Å
Structure data

SASDAT6:
Integrin beta4, fragment of the cytoplasmic region encompassing the third and fourth FnIII domains
Method: SAXS/SANS

PDB-4wtw:
Crystal structure of the third FnIII domain of integrin beta4
Method: X-RAY DIFFRACTION / Resolution: 1.606 Å

PDB-4wtx:
Crystal structure of the fourth FnIII domain of integrin beta4
Method: X-RAY DIFFRACTION / Resolution: 1.5 Å

Chemicals

ChemComp-SO4:
SULFATE ION

ChemComp-1PE:
PENTAETHYLENE GLYCOL / precipitant*YM

ChemComp-EDO:
1,2-ETHANEDIOL

ChemComp-HOH:
WATER

Source
  • homo sapiens (human)
KeywordsCELL ADHESION / IMMUNOGLOBULIN FOLD / FIBRONECTIN TYPE III / INTEGRIN / RECEPTOR

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