Abstract
Radiollgand binding is a straightforward technique that measures the bindmg of a labeled agonist or antagonist to its receptor. It is applicable to a variety of receptor preparations, ranging from purified receptors to tissue slices, or even whole animals. However, membranes or broken-cell preparations are undoubtedly the most widely used. Radioligand binding allows the affinity of drugs for their receptors to be determined very readily and It also allows the number of receptors in a tissue or cell to be quantified-something that was impossible before the mtroductlon of binding techniques. Furthermore, the technique can be adapted to study the association and dissociation kinetics of llgand binding (1,2), as well as complex allosteric mteractlons between ligands (1) or between receptors and effector molecules, such as guanine nucleotide-binding proteins (G proteins) (1,2).
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References
Hulme E. C., ed. (1992) Receptor-Ligand Znteractzons. A Practical Approach. IRL Press at Oxford University Press, Oxford, UK.
Keen M. and MacDermot J. (1993) Analysis of receptors by radioligand binding, in Receptor Autoradiography: Principles and Practice (Wharton J. and Polak J. M., eds.), Oxford University Press, Oxford, UK, pp. 23–56.
Keen M. (1995) The problems and pitfalls of radioligand binding, in Methods in Molecular Bzology, vol. 41. Szgnal Transduction Protocols (Kendall D. A. and Hill S. J., eds.), Humana, Totowa, NJ, pp. 1–16.
Klotz I. M. (1982) Numbers of receptor sites from Scatchard graphs: facts and fantasies. Science 217, 1247–1249.
Burgisser E. (1984) Radioligand-receptor binding studies: What’s wrong with the Scatchard analysis? Trends Pharmacol. Sci. 5, 142–144.
Cheng Y. C. and Prusoff W. H. (1973) Relationship between the inhibition constant Kl and the concentration of inhibitor which causes 50% inhibition (IC50) of an enzymic reaction. Biochem. Pharmacol. 22, 3099–3108.
Kelly E., Keen M., Nobbs P., and MacDermot J. (1990) Segregation of discrete G, sα-mediated responses that accompany homologous or heterologous desensitization in two related somatic hybrids. Br J. Pharmacol. 99, 309–316.
Nathanson N. M. (1983) Binding of agonists and antagonists to muscarimc acetylcholine receptors on intact cultured heart cells. J. Neurochem. 41, 1545–1549.
Sibley D. R. and Lefkowitz R. J. (1985) Molecular mechanisms of receptor desensitization using the P-adrenergic receptor-coupled adenylate cyclase system as a model. Nature 317, 124–129.
Wise H. and Jones R. L. (1996) Focus on prostacyclin and its novel mimetics. Trends Pharmacol. Sci. 17, 17–21.
MacDermot J., Blair I., and Cresp T. M. (1981) Prostacyclin receptors of a neuronal hybrid cell line; divalent cations and ligand-receptor coupling. Biochem. Pharmacol. 30, 2041–2044.
Motulsky H. J. and Mahan L. C. (1984) The kinetics of competitive radioligand binding predicted by the law of mass action. Mol. Pharmacol. 25, 1–9.
Tobler H. J. and Engel G. (1983) Affinity spectra: a novel way for the evaluation of equilibrium binding experiments. Naunyn-Schmiedeberg’s Arch. Pharmacol. 322, 183–192.
Stockton J. M., Birdsall N. J. M., Burgen A. S. V., and Hulme E. C. (1983) Modification of the binding properties of muscarinic acetylcholine receptors by gallamine. Mol Pharmacol. 23, 55l–557.
Wells J. W., Birdsall N. J. M., Burgen A. S. V., and Hulme E. C. (1980) Competitive binding studies with multiple sites: effects arising from depletion of free radioligand. Biochim. Btophys Acta 632, 464–469.
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Keen, M. (1997). Radioligand-Binding Methods for Membrane Preparations and Intact Cells. In: Challiss, R.A.J. (eds) Receptor Signal Transduction Protocols. Methods in Molecular Biology™, vol 83. Humana Press. https://doi.org/10.1385/0-89603-495-X:1
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DOI: https://doi.org/10.1385/0-89603-495-X:1
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