Temperature-induced unfolding of ribonuclease A embedded in spherical polyelectrolyte brushes.
We use Fourier Transform infrared spectroscopy (FT-IR) spectroscopy to study the thermal unfolding and refolding behavior of ribonuclease (RNase A) adsorbed to spherical polyelectrolyte brushes (SPB). The SPB consist of a solid poly(styrene) core of ca. 100 nm diameter onto which long chains of poly(styrene sulfonic acid), PSS have been densely attached. The particles bearing the adsorbed protein are dispersed in aqueous buffer solution at a pH close to the isoelectric point (9.6) of the protein. The secondary structure of the protein was analyzed by FT-IR spectroscopy and compared to the structure of the native protein before adsorption. The unfolding of the free RNase A in solution was found to be fully reversible with an unfolding temperature of 65 degrees C, in accordance to previous studies. However, after adsorption to the SPB, the unfolding temperature of the protein molecule is lowered by 10 degrees C and the Van't Hoff enthalpy of the unfolding process is significantly reduced. Moreover the unfolding of the adsorbed protein is irreversible. The phenomenon may be explained by an increase in binding sites due to unfolding of the globular structure. Protein adsorption to a spherical polyelectrolyte brush.
Wittemann
Alexander
A
Physikalische Chemie I, Universität Bayreuth, Universitätsstrasse 30, 95440 Bayreuth, Germany. Alexander.Wittemann@uni-bayreuth.de
Ballauff
Matthias
M
D000327:Adsorption; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D004573:Electrolytes / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008954:Models, Biological; D008958:Models, Molecular; D011137:Polystyrenes / Q000737:chemistry; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry; D012259:Ribonuclease, Pancreatic / Q000737:chemistry*; D012542:Scattering, Radiation; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D013816:Thermodynamics; D014965:X-Rays
D004573:Electrolytes; D011137:Polystyrenes; D011506:Proteins; C003321:polystyrene sulfonic acid; D012259:Ribonuclease, Pancreatic