Oligomerisation and thermal stability of polyvalent integrin α5β1 ligands

Michaela Kreiner, Olwyn Byron, Diana Domingues, Christopher F. van der Walle

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Synthetic oligomeric integrin α5β1 ligands, specifically immobilised to surfaces, facilitate increased fibroblast cell spreading compared with that associated with the monomer. These ligands consist of a N-terminal fibronectin domain pair, a spacer and a di-, tri- or tetrameric coiled coil. However, it is not yet clear what effect fusion of the fibronectin domains has on the predicted oligomerisation of the coiled coils. Using analytical ultracentrifugation we show that the predicted tetrameric and trimeric coiled coils facilitate a corresponding ligand oligomerisation with half-dissociation at 0.7 and 0.2 µM, respectively. In contrast, the predicted dimeric coiled coil formed both dimers and trimers. Under non-reducing conditions, the unique C-terminal thiol-facilitated inter-oligomer dimerisation of the trimeric species, generating hexameric ligands. Disulphide bonding also increased helical stability during thermal unfolding. The work allows the cellular response to these clustered integrin α5β1 ligands to be more accurately interpreted, and has wider implications with respect to the utility of coiled coils as tools to facilitate protein oligomerisation.
LanguageEnglish
Pages34-39
Number of pages5
JournalBiophysical Chemistry
Volume142
Issue number1-3
DOIs
Publication statusPublished - 2009

Fingerprint

Oligomerization
Integrins
Thermodynamic stability
Hot Temperature
Ligands
Fibronectins
Dimerization
Ultracentrifugation
Fibroblasts
Oligomers
Sulfhydryl Compounds
Disulfides
Dimers
Fusion reactions
Monomers
Cells
Proteins

Keywords

  • coiled coil
  • fibronectin
  • integrin
  • analytical ultracentrifugation
  • thermal unfolding

Cite this

Kreiner, Michaela ; Byron, Olwyn ; Domingues, Diana ; van der Walle, Christopher F. / Oligomerisation and thermal stability of polyvalent integrin α5β1 ligands. In: Biophysical Chemistry. 2009 ; Vol. 142, No. 1-3. pp. 34-39.
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Oligomerisation and thermal stability of polyvalent integrin α5β1 ligands. / Kreiner, Michaela; Byron, Olwyn; Domingues, Diana; van der Walle, Christopher F.

In: Biophysical Chemistry, Vol. 142, No. 1-3, 2009, p. 34-39.

Research output: Contribution to journalArticle

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