title: Effects of thermodynamic nonideality in ligand binding studies.
abstract: Effects of thermodynamic nonideality are considered in relation to the quantitative characterization of the interaction between a small ligand. S, and a macromolecular acceptor. A, by two types of experimental procedure. The first involves determination of the concentration of ligand in dialysis equilibrium with the acceptor/ligand mixture, and the second, measurement of the concentration of unbound ligand in the reaction mixture by ultrafiltration or the rate of dialysis method. For each situation explicit expressions are formulated for the appropriate binding function with allowance for composition-dependent nonideality effects expressed in terms of molar volume, charge-charge interaction and covolume contributions. The magnitudes of these effects are explored with the aid of experimental studies on the binding of tryptophan and of methyl orange to bovine serum albumin. It is concluded for experiments conducted utilizing either equilibrium dialysis or frontal gel chromatography that, provided a correction is made for any Donnan redistribution of ligand, theoretically predicted acceptor-concentration dependence is likely to be negligible and that use of the conventional binding equation written for an ideal system is appropriate to the analysis of the results. Use of ultrafiltration or the rate of dialysis method requires examination of the assumption that the activity coefficient ratio y(A)y(s)/y(AS) for the reaction mixture approximates unity; but again reassurance is provided that nonideality manifested as a dependence of the binding function on acceptor concentration is unlikely to be significant.
authors.affiliation: Department of Biochemistry, University of Queensland, St Lucia, Queensland 4067, Australia.
authors.forename: C L
authors.initials: CL
authors.lastname: Ford
authors.forename: D J
authors.initials: DJ
authors.lastname: Winzor
authors.forename: L W
authors.initials: LW
authors.lastname: Nichol
authors.forename: M J
authors.initials: MJ
authors.lastname: Sculley