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Binding issues. A scientist wishes to determine the number of receptors specific for a ligand $mathrm{X}$, which he has in both radioactive and nonradioactive form. In one experiment, he adds increasing amounts of the radioactive $mathrm{X}$ and measures how much of it is bound to the cells. The result is shown as total activity in the graph. Next, he performs the same experiment, except that he includes a several hundredfold excess of nonradioactive X. This result is shown as nonspecific binding. The difference between the two curves is the specific binding. (a) Why is the total binding not an accurate representation of the number of receptors on the cell surface? (b) What is the purpose of performing the experiment in the presence of excess nonradioactive ligand? (c) What is the significance of the fact that specific binding attains a plateau? 15. Counting receptors. With the use of experiments such as those described in problems 13 and 14, it is possible to calculate the number of receptors in the cell membrane. Suppose that the specific activity of the ligand is $10^{12}$ cpm per millimole and that the maximal specific binding is $10^{4}$ cpm per milligram of membrane protein. There are $10^{10}$ cells per milligram of membrane protein. Assume that one ligand binds per receptor. Calculate the number of receptor molecules present per cell.
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