โ Scribed by Jean-Pierre Raynaud
No coin nor oath required. For personal study only.
This program has been developed for the estimation and statistical evaluation of binding parameters. Binding of small molecules to protein mixtures is measured at equilibrium and the results analyzed on the assumption that each binding system is independent and follows the law of mass action. Briefly, the method consists in plotting the log of the fraction bound or unbound against the log of the total ligand concentration in a Proportion Graph [ 1 ] and estimating the binding parameters first by graphical construction, then by multiple regression analysis. The estimates thus obtained are then improved by a least squares method. Finally, a statistical check is carried out to verify whether the experimental results fit the chosen model well.
The method affords the distinct advantage of enabling a composite non-linear curve to be broken down into its constituent elements, thus yielding much more detailed information than the classical linear Lineweaver-Burk [2] and Eadie-Scatchard plots [3]. Examples are given of the application of the program to the analysis of mixtures involving two kinds of binding: specific binding (high affinity, low capacity, high ligand specificity) and non-specific binding (low affinity, high capacity, no ligand specificity).
The computer print-out not only gives the binding parameter values, but also a graphical representation of the experimental and calculated binding curve in a Proportion Graph thus illustrating the adequacy of fit..
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