title: On the need to consider kinetic as well as thermodynamic consequences of the parking problem in quantitative studies of nonspecific binding between proteins and linear polymer chains.
abstract: Attention is drawn to a need for caution in the thermodynamic characterization of nonspecific binding of a large ligand to a linear acceptor such as a polynucleotide or a polysaccharide-because of the potential for misidentification of a transient (pseudoequilibrium) state as true equilibrium. The time course of equilibrium attainment during the binding of a large ligand to nonspecific three-residue sequences of a linear acceptor lattice has been simulated, either by numerical integration of the system of ordinary differential equations or by a Monte Carlo procedure, to identify the circumstances under which the kinetics of elimination of suboptimal ligand attachment (called the parking problem) create such difficulties. These simulations have demonstrated that the potential for the existence of a transient plateau in the time course of equilibrium attainment increases greatly (i) with increasing extent of acceptor saturation (i.e., with increasing ligand concentration), (ii) with increasing magnitude of the binding constant, and (iii) with increasing length of the acceptor lattice. Because the capacity of the polymer lattice for ligand is most readily determined under conditions conducive to essentially stoichiometric interaction, the parameter so obtained is thus likely to reflect the transient (irreversible) rather than equilibrium binding capacity. A procedure is described for evaluating the equilibrium capacity from that irreversible parameter; and illustrated by application to published results [M. Nesheim, M.N. Blackburn, C.M. Lawler, K.G. Mann, J. Biol. Chem. 261 (1986) 3214-3221] for the stoichiometric titration of heparin with thrombin.
authors.affiliation: Centre for Protein Structure, Function and Engineering, Department of Biochemistry, University of Queensland, Brisbane, Queensland 4072, Australia.
authors.forename: P D
authors.initials: PD
authors.lastname: Munro
authors.forename: C M
authors.initials: CM
authors.lastname: Jackson
authors.forename: D J
authors.initials: DJ
authors.lastname: Winzor