title: Analysis of protein self-association under conditions of the thermodynamic non-ideality.
abstract: Analysis of protein-protein interactions in highly concentrated solutions requires a consideration of the non-ideality in such solutions which is expressed by the virial coefficients. Different equations are presented to estimate effects of the thermodynamic non-ideality on the macromolecular interaction of self-associating proteins in sedimentation equilibrium experiments. Usually the influence of thermodynamic non-ideal behavior are described by concentration power series. The convergence of such power series is limited at high solute concentration. When expressing the thermodynamic non-ideality by an activity power series this disadvantage can be minimized. The developed centrifuge equations are the basis for a global analysis to estimate equilibrium constants and the corresponding thermodynamic activities of the reactants. Based on fit analysis of synthetic concentration profiles it was established that marked deviations from the expected association constants are observed for proteins with strong association forces between solute molecules. Considerable differences were also observed in weakly interacting systems. This was due to the excluded volume of the protein which is similar in magnitude to the binding constant. For interactions with moderate affinities values extremely close to the true binding values were obtained, as confirmed by experimental results with concanavalin A.
authors.affiliation: Max Delbrück Center for Molecular Medicine, Berlin, Germany. behlke@mdc-berlin.de
authors.forename: J
authors.initials: J
authors.lastname: Behlke
authors.forename: O
authors.initials: O
authors.lastname: Ristau
mesh_terms: D003208:Concanavalin A / Q000737:chemistry* / Q000378:metabolism; D046911:Macromolecular Substances; D008956:Models, Chemical; D011485:Protein Binding; D012997:Solvents; D013816:Thermodynamics; D014461:Ultracentrifugation
chemical_list: D046911:Macromolecular Substances; D012997:Solvents; D003208:Concanavalin A