Biochemistry
Biochemistry
9th Edition
ISBN: 9781319114671
Author: Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher: W. H. Freeman
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**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.

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**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?

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**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.
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Transcribed Image Text: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.
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