Complex Binding Phenomena

  • Lee E. Limbird


The methods for acquisition and initial analysis of radioligand binding phenomena were summarized in chapter 3. It was demonstrated that equations for linear transformations of binding data were derived assuming that a reversible bimolecular reaction occurred between ligand and receptor and that this interaction obeyed mass action law, namely *D + R ⇌ *DR. Consequently, when data transformations such as the Scatchard plot are nonlinear, when Hill coefficients (nH) do not equal 1.0, or when competition binding curves are not of normal steepness, additional complexities are suggested. Chapter 3 also provided guidelines for evaluating whether or not technical artifacts were responsible for departure of the data from that expected for a simple bimolecular reaction. Once technical artifacts have been excluded, complex binding phenomena suggest the existence of biological complexities which may provide insights into the molecular basis of receptor function.


Receptor Subtype Guanine Nucleotide Slope Factor Agonist Binding Receptor Population 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.



  1. DeLean, A., Hancock, A.A. and Lefkowitz, R.J. (1981) Validation and statistical analysis of a computer modeling method for quantitative analysis of radioligand binding data for mixtures of pharmacological receptor subtypes. Mol. Pharmacol. 21:5–16.Google Scholar
  2. DeLean, A., Munson, P.J. and Rodbard, D. (1978) Simultaneous analysis of families of sigmoidal curves: Application to bioassay, radioligand assay and physiological dose-response curves. Am. J. Physiol. 235:E97–E102.PubMedGoogle Scholar
  3. DeLean, A., Stadel, J.M. and Lefkowitz, R.J. (1980) A ternary complex model explains the agonist-specific binding properties of the adenylate cyclase-coupled β-adrenergic receptor. J. Biol. Chem. 255:7108–7117.Google Scholar
  4. Janin, J. (1973) The study of allosteric proteins. Prog. Biophys. Mol. Biol. 27:77–119.CrossRefGoogle Scholar
  5. Klotz, I.M. (1946) The application of the law of mass action to binding by proteins. Interactions with calcium. J. Am. Chem. Soc. 9:109–117.Google Scholar
  6. Klotz, I.M. and Hunston, D.L. (1975) Protein interactions with small molecules: Relationships between stoichimetric binding constants, site binding constants, and empirical binding parameters. J. Biol. Chem. 250:3001–3009.PubMedGoogle Scholar
  7. Koshland, D.E., Nemethy, G. and Filmer, D. (1966) Comparison of experimental binding data and theoretical models in proteins containing subunits. Biochem. 5:365–385.CrossRefGoogle Scholar
  8. Molinoff, P.B., Wolfe, B.B. and Weiland, G.A. (1981) Quantitative analysis of drug-receptor interactions II. Determination of the properties of receptor subtypes. Life Sci. 29:427–443.PubMedCrossRefGoogle Scholar
  9. Munson, P.J. (1983) LIGAND: A computerized analysis of ligand binding data. Methods in Enzymology 92:543–546.PubMedCrossRefGoogle Scholar
  10. Newsholme, E.A. and C. Start (1973) Regulation in Metabolism, (ed.), ch. 2. New York: John Wiley and Sons.Google Scholar
  11. Steinhardt, J. and Reynolds, J.A. (1969) Multiple Equilibria in Proteins, (ed.), ch. 2, pp. 10–33. New York: Academic Press.Google Scholar
  12. Teipel, J. and Koshland, D.E. (1969) The significance of intermediary plateau regions in enzyme saturation curves. Biochem. 8:4656–4663.CrossRefGoogle Scholar
  13. Wregett, K.A. and DeLean, A. (1984) The ternary complex model. Its properties and application to ligand interactions with the D2-dopamine receptor of the anterior pituitary gland. Mol. Pharmacol. 26:214–227.Google Scholar


  1. Adair, G.S. (1925) The hemoglobin system. VI. The oxygen dissociation curve of hemoglobin. J. Biol Chem. 63:529–545.Google Scholar
  2. Barnett, D.B., Rugg, E.L. and Nahorski, S.R. (1978) Direct evidence of two types of β-adrenoceptor binding sites in lung tissue. Nature 273:166–168.PubMedCrossRefGoogle Scholar
  3. Birdsall, N.J.M., Hulme, E.C. and Burgen, A.S.V. (1980). The character of the muscarinic receptors in different regions of the rat brain. Proc. Roy. Soc. Lond. B. 207:1–12.CrossRefGoogle Scholar
  4. Burgisser, E., DeLean, A. and Lefkowitz, R.J. (1982) Reciprocal modulation of agonistand antagonist binding to muscarinic cholinergic receptors by guanine nucleotide. Proc. Natl. Acad. Sci. USA 79:1732–1736.PubMedCrossRefGoogle Scholar
  5. DeHaen, C. (1976) The non-stoichiometric floating receptor model for hormone sensitive adenylyl cyclase. J. Theoret. Biol. 58:383–400.CrossRefGoogle Scholar
  6. Feldman, H.A. (1972) Mathematical theory of complex ligand-binding systems at equilibrium: Some methods for parameter fitting. Anal. Biochem. 48:317–338.PubMedCrossRefGoogle Scholar
  7. Hoffman, B.B., DeLean, A., Wood, C.L., Schocken, D.D. and Lefkowitz, R.J. (1979) Alphaadrenergic receptor subtypes: Quantitative assessment by ligand binding. Life Sci. 24:1739–1746.PubMedCrossRefGoogle Scholar
  8. Hoffman, B.B. and Lefkowitz, R.J. (1980) An assay for alpha-adrenergic receptor subtypes using [3H]-dihydroergocryptine. Biochem. Pharmacol. 29:452–454.PubMedCrossRefGoogle Scholar
  9. Jacobs, S. and Cuatrecasas, P. (1976) The mobile receptor hypothesis and “cooperativity” of hormone binding. Application to insulin. Biochim. Biophys. Acta 433:482–495.PubMedCrossRefGoogle Scholar
  10. Katz, B. and Thesleff, S. (1957) A study of the “desensitization” produced by acetylcholine at the motor end plate. J. Physiol. 138:63–80.PubMedGoogle Scholar
  11. Kent, R.S., DeLean, A. and Lefkowitz, R.J. (1980) A quantitative analysis of beta-adrenergic receptor interactions: Resolution of high and low affinity states of the receptor by computer modeling of ligand binding data. Mol. Pharmacol. 17:14–23.PubMedGoogle Scholar
  12. Kilpatrick, B.V. and Caron, M.G. (1983) Agonist binding promotes a guanine nucleotide reversible increase in the apparent size of the bovine anterior pituitary dopamine receptors. J. Biol. Chem. 258:13528–13534.PubMedGoogle Scholar
  13. Klotz, I.M. (1983) Ligand-receptor interactions: What we can and cannot learn from binding measurements. Trends in Pharmacol. Sci. 4:253–255.CrossRefGoogle Scholar
  14. Klotz, I.M. and Hunston, D.L. (1984) Mathematical models for ligand-receptor binding. Real sites, ghost sites. J. Biol. Chem. 259:10060–10062.PubMedGoogle Scholar
  15. Lavin, T.N., Hoffman, B.B. and Lefkowitz, R.J. (1981) Determination of subtype selectivity of alpha-adrenergic ligands. Comparison of selective and non-selective radioligands. Mol. Pharmacol. 20:28–34.PubMedGoogle Scholar
  16. Limbird, L.E., Gill, D.M. and Lefkowitz, R.J. (1980) Agonist-promoted coupling of the β-adrenergic receptor with the guanine nucleotide regulatory protein of the adenylate cyclase system. Proc. Natl. Acad. Sci. USA 77:775–779.PubMedCrossRefGoogle Scholar
  17. Michel, T.M., Hoffman, B.B., Lefkowitz, R.J. and Caron, M.G. (1981) Different sedimentation properties of agonist- and antagonist-labeled platelet alpha2-adrenergic receptors. Biochem. Biophys. Res. Commun. 100:1131–1134.PubMedCrossRefGoogle Scholar
  18. Minneman, K.P., Hegstrand, L.R. and Molinoff, P.B. (1979) Simultaneous determination of beta1 and beta2-adrenergic receptors in tissues containing both subtypes. Mol. Pharmacol. 16:34–46.PubMedGoogle Scholar
  19. Munson, P.J. and Rodbard, D. (1980) LIGAND: A versatile computerized approach for characterization of ligand-binding systems. Anal. Biochem. 107:220–239.PubMedCrossRefGoogle Scholar
  20. Rugg, E.L., Barnett, D.L. and Nahorski, S.R. (1978) Coexistence of beta1 and beta2 adrenoceptors in mammalian lung: evidence from direct binding studies. Mol. Pharm. 14:996–1005.Google Scholar
  21. Smith, S.K. and Limbird, L.E. (1981) Solubilization of human plateletα -adrenergic receptors: Evidence that agonist occupancy of the receptor stabilizes receptor-effector interactions. Proc. Natl. Acad. Sci. USA 78:4026–4030.PubMedCrossRefGoogle Scholar
  22. Weiland, G.A., Minneman, K.P. and Molinoff, P.B. (1979) Fundamental difference between the molecular interactions of agonists and antagonists with the β-adrenergic receptor. Nature 281:114–117.PubMedCrossRefGoogle Scholar
  23. Williams, L.T. and Lefkowitz, R.J. (1977) Slowly reversible binding of catecholamine to a nucleotide-sensitive state of the β-adrenergic receptor. J. Biol. Chem. 252:7207–7213.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 1986

Authors and Affiliations

  • Lee E. Limbird
    • 1
  1. 1.Vanderbilt UniversityNashvilleUSA

Personalised recommendations