Here is the transcribed content from the image suitable for an educational website:---**Equilibrium Dialysis and cAMP Binding Protein Analysis**A cyclic AMP (cAMP) binding protein was isolated from mammalian cells and characterized using equilibrium dialysis experiments. The data from these experiments is provided below. The protein was present in the dialysis bags at a concentration of 16 μM.---**Concentration of cAMP (μM)**| Outside Bag | Inside Bag ||-------------|------------|| 0.7 | 2.7 || 2.3 | 8.3 || 4.5 | 15.0 || 10.0 | 26.0 || 30.0 | 54.0 || 70.0 | 98.0 || 150.0 | 180.0 |---**Analysis Questions:**i) Using a Scatchard plot, estimate the Kd for the cAMP.ii) Estimate the number of binding sites per protein molecule.iii) What other information could be obtained from this analysis?---**Instructions for Further Analysis:**To analyze the binding characteristics of the cAMP-binding protein, you would typically use a Scatchard plot to determine the dissociation constant (Kd) and the number of binding sites. Plot the ratio of bound to free ligand versus the concentration of bound ligand. This analysis will provide insights into the affinity and capacity of the protein for cAMP, valuable for understanding its functional role in cellular processes.

Given that,The ligand is cAMPThe receptor is cAMP binding protein (CBP)also the concentration of CBP is 16 μM inside the dialysis bagthe concentration of ligand is given in the table below.Scatchard analysis involves plotting bound ligand [LR]ligand [L] vs LR on a graphthe slope gives us -1KD dissociation constant and the X intercept gives number of binding sites of the receptor outside bag (free cAMP) inside bag (free cAMP+ bound cAMP) bound cAMP (inside - outside) bound cAMP/ free cAMP 0.7 2.7 2 3.8 2.3 8.3 6 2.6 4.5 15 10.5 2.33 10 26 16 1.6 30 54 24 0.8 70 98 28 0.4 150 180 30 0.2the line equation for the plot is : y=-0.118×RL+3.633looks like y=mx+chence the slope is m=-0.118we know,slope=-1KD-0.118=-1KDKD=10.118=8.47 μMUsing the equation,y=-0.118×RL+3.633we found the KD value of cAMPnow, from the Scatchard analysis, we know that the X intercept gives the number of binding sites of the receptor.X intercept value can be determined from the line equation itself. The value of X when Y is 0 is called as X intercept.0=-0.118×RL+3.633RL=3.6330.118≈31μMnow, given that the concentration of free CBP is 16 μM inside the bag,Then RLR=3116≈2It means there is twice the amount of bound receptor than free receptor. Therefore, the receptor has 2 binding sites for the ligand.From the table we can see that the concentration of the bound cAMP is far less than the concentration of free cAMP. This is evident from the low KD value of cAMP. This is essential as cAMP mediates a number of processes downstream of a signal cascade. If the association is tight, the signal would persist longer than it would need to be.