Interdomain mobility and conformational stability of type III fibronectin domain pairs control surface adsorption, desorption and unfolding

Pereira, P., Kelly, S.M. , Gellert, P.R. and van der Walle, C.F. (2008) Interdomain mobility and conformational stability of type III fibronectin domain pairs control surface adsorption, desorption and unfolding. Colloids and Surfaces B: Biointerfaces, 64(1), pp. 1-9. (doi:10.1016/j.colsurfb.2007.12.015)

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Publisher's URL: http://dx.doi.org/10.1016/j.colsurfb.2007.12.015

Abstract

The 9th-10th type III fibronectin domain pair (9-10FNIII) has found widespread use as a biomimetic surface for cell adhesion. However, the effect of mutations to 9-10FNIII on its surface adsorption characteristics have not been investigated. Here we address this issue using total internal reflection fluorescence (TIRF) and circular dichroism spectroscopy, comparing two conformationally stable 9-10FNIII mutants against the wild type. Desorption of the 9-10FNIII mutants from the silica surface was minimal in comparison to desorption of 9-10FNIII. The extent and rate of protein desorption from silica was empirically matched by loss of secondary structure upon adsorption, with only the spectrum for 9-10FNIII showing extensive loss of the [beta]-sandwich fold. For the proteins adsorbed to hydrophobic surfaces, only the CD spectra for the 9-10FNIII mutant constrained via an interdomain disulphide bridge showed similarity with the corresponding solution structure. Since the binding of 9-10FNIII to integrin [alpha]5[beta]1 is highly dependent on the relative spatial arrangement of the two domains, we suggest that the observed differences in cell adhesion and spreading on wild type 9-10FNIII and mutants may in part be attributed to the extent of protein desorption and unfolding at the surface

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Kelly, Dr Sharon
Authors: Pereira, P., Kelly, S.M., Gellert, P.R., and van der Walle, C.F.
Subjects:Q Science > QH Natural history > QH345 Biochemistry
College/School:College of Medical Veterinary and Life Sciences
Journal Name:Colloids and Surfaces B: Biointerfaces
ISSN:0927-7765

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