Determination of the parameters of self-association by direct fitting of the omega function.
Nonlinear regression is used to fit the omega function vs. protein concentration curves (first described by B.K. Milthorpe, P.D. Jeffrey and L.W. Nichol, Biophys. Chem. 3 (1975) 169) obtained from sedimentation equilibrium experiments on self-associating macromolecules. Nonlinear regression allows the direct fit of these curves with discrete or indefinite self-association reaction models in order to obtain values for the equilibrium constants and second virial coefficient. The method is independent of the choice of reference concentration and avoids the original method of extrapolating an omega function curve to zero concentration and then using the extrapolated value to construct a monomer activity curve used for analysis. This extrapolation can become very difficult for mild to strong self-associations where incorrectly extrapolated values lead to systematic error in the monomer activity curves. The method is applied to results from a mild, indefinite self-association, exemplified by the self-association of human spectrin, and to computer-simulated data of weak, mild and strong, indefinite self-associations.
D004912:Erythrocytes / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D008433:Mathematics; D011506:Proteins / Q000378:metabolism*; D013049:Spectrin / Q000378:metabolism*
D046911:Macromolecular Substances; D011506:Proteins; D013049:Spectrin