Kinetics of conformational changes of fibronectin adsorbed onto model surfaces.
Fibronectin (FN), a large glycoprotein found in body fluids and in the extracellular matrix, plays a key role in numerous cellular behaviours. We investigate FN adsorption onto hydrophilic bare silica and hydrophobic polystyrene (PS) surfaces using Fourier transform infrared spectroscopy-attenuated total reflection (FTIR-ATR) in aqueous medium. Adsorption kinetics using different bulk concentrations of FN were followed for 2h and the surface density of adsorbed FN and its time-dependent conformational changes were determined. When adsorption occurs onto the hydrophilic surface, FN molecules keep their native conformation independent of the adsorption conditions, but the amount of adsorbed FN increases with time and the bulk concentration. Although the protein surface density is the same on the hydrophobic PS surface, this has a strong impact on the average conformation of the adsorbed FN layer. Indeed, interfacial hydration changes induced by adsorption onto the hydrophobic surface lead to a decrease in unhydrated beta-sheet content and cause an increase in hydrated beta-strand and hydrated random domain content of adsorbed FN. This conformational change is mainly dependent on the bulk concentration. Indeed, at low bulk concentrations, the secondary structures of adsorbed FN molecules undergo strong unfolding, allowing an extended and hydrated conformation of the protein. At high bulk concentrations, the molecular packing reduces the unfolding of the stereoregular structures of the FN molecules, preventing stronger spreading of the protein.
Baujard-Lamotte
L
L
Laboratoire ERRMECe, Université de Cergy-Pontoise, 95302 Cergy-Pontoise Cedex, France. lucie.baujard@u-cergy.fr <lucie.baujard@u-cergy.fr>
Noinville
S
S
Goubard
F
F
Marque
P
P
Pauthe
E
E
D000327:Adsorption; D005353:Fibronectins / Q000737:chemistry*; D007202:Indicators and Reagents; D007700:Kinetics; D008956:Models, Chemical; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D012822:Silicon Dioxide; D017550:Spectroscopy, Fourier Transform Infrared; D013499:Surface Properties
D005353:Fibronectins; D007202:Indicators and Reagents; D012822:Silicon Dioxide