Analyzing Radioligand Binding Data

Harvey Motulsky1, Richard Neubig2

1 GraphPad Software, San Diego, 2 University of Michigan, Ann Arbor
Publication Name:  Current Protocols in Protein Science
Unit Number:  Appendix 3H
DOI:  10.1002/0471140864.psa03hs21
Online Posting Date:  May, 2001
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Abstract

A radioligand is a radioactively labeled drug that can associate with a receptor, transporter, enzyme, or any protein of interest. Measuring the rate and extent of binding provides information on the number of binding sites, and their affinity and accessibility for various drugs. Radioligand binding experiments are easy to perform, and provide useful data in many fields. For example, radioligand binding studies are used to study receptor regulation, investigate receptor localization in different organs or regions using autoradiography, categorize receptor subtypes, and probe mechanisms of receptor signaling. This unit reviews the theory of receptor binding and explains how to analyze experimental data. Since binding data are usually best analyzed using nonlinear regression, this unit also explains the principles of curve fitting with nonlinear regression.

     
 
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Table of Contents

  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

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Figures

Videos

Literature Cited

Literature Cited
   Cheng, Y. and Prusoff, W.H. 1973. Relationship between the inhibition constant (Ki) and the concentration of an inhibitor that causes a 50% inhibition (I50) of an enzymatic reaction. Biochem. Pharmacol. 22:3099‐3108.
   Kenakin, T. 1993. Pharmacologic Analysis of Drug‐Receptor Interactions. Raven Press, New York.
   Limbird, L.E. 1996. Cell Surface Receptors: A Short Course in Theory and Methods, 2nd ed. Kluwer Academic Publishers, Boston.
   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.
   Munson, P.J. and Rodbard, D. 1980. Ligand: A versatile computerized approach to characterization of ligand binding systems. Anal. Biochem. 107:220‐239.
   Rosenthal, H.E. 1967. A graphic method for the determination and presentation of binding parameters in complex systems. Anal. Biochem. 20:525‐532.
   Swillens, S. 1995. Interpretation of binding curves obtained with high receptor concentrations: Practical aid for computer analysis. Mol. Pharmacol. 47:1197‐1203.
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