Binding Constants: Measurement and Biological Range


Several methods exist for the measurement of binding constants for macromolecular interactions between dissimilar reactants. These allow biological interactions with binding constants in the range 102 to 109 mol L−1 to be characterized quantitatively.

Keywords: equilibrium interactions; reaction thermodynamics; ligand–acceptor (receptor) interactions

Figure 1.

Presentation of data for the interaction of a univalent ligand with acceptor. (a) Simulated rectangular hyperbolic dependence of the binding function upon free ligand concentration (eqn ) for an acceptor with four sites (n = 4) governed by an intrinsic binding constant (KAS) of 105 mol L−1. (b) Scatchard linear transformation (eqn ) of the same dependence.

Figure 2.

Allowance for ligand multivalence in the analysis of binding data. (a) Multivalent equivalent of the rectangular hyperbolic dependence of binding function upon free ligand concentration (eqn ), simulated for a system with f = 2, n = 1, KAS = 105 mol L−1 and [A]tot = 20 μmol L−1. (b) General counterpart of the Scatchard plot (eqn ) of the same dependence. (c) Conventional Scatchard plot of the same data.

Figure 3.

Evaluation of binding constants by biosensor technology. Simulated time dependence of biosensor response resulting from injection (at 5 μL min−1) of 30 μL of A (100 nmol L−1) followed by buffer, B, (eqns and ) for a system with Rmax = 3000 units, ka = 105 mol L−1 s−1 and kd = 10−2 s−1. 1000 B/A core units = 0.1 degree. Inset: determination of the association and dissociation rate constants from the dependence of the pseudo‐first‐order rate constant (kobs) upon the injected concentration of A (eqn ).



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Further Reading

Cserháti T and Valkó K (1994) Chromatographic Determination of Molecular Interactions. Boca Raton, FL: CRC Press.

Hall DR and Winzor DJ (1998) Potential of biosensor technology for the characterization of interactions by quantitative affinity chromatography. Journal of Chromatography B 715: 163–181.

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Lundahl P, Greiger E and Lundqvist A (eds) (1998) Quantitative Analysis of Biospecific Interactions. Chur, Switzerland: Harwood.

Winzor DJ (1997) Quantitative affinity chromatography. In: Matetschuk P (ed.) Affinity Separations: A Practical Approach, pp. 39–60. Oxford: IRL Press.

Winzor DJ and Sawyer WH (1995) Quantitative Characterization of Ligand Binding. New York: Wiley–Liss.

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Winzor, Donald J(Apr 2001) Binding Constants: Measurement and Biological Range. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1038/npg.els.0000615]