5:36 PM c 56%. old examsR 2016 3510_Final.pdf 11) (4 points) Ligand binding curv

Question

5:36 PM c 56%. old examsR 2016 3510_Final.pdf 11) (4 points) Ligand binding curves for binding of two ligands, HABA and 2-Iminobiotin (2IMB), to egg white avidin are shown on the right 1.0 0.8 What is the approximate Kd for HABA binding to avidin, 0.0 0.4 B) 10 uM 02 D) 100 M 10 20 30 Ligand] (uM Based on the two plots, which ligand binds more tightly to avidin, HABA or 2IMB? A) HABA B) 2-iminibiotin 12) (2 points) In a mixture of the five proteins listed below, which should elute second in size- exclusion (gel filtration) chromatography A) cytochrome c B) immunoglobulin G M 145,000 C) ribonuclease A D) RNA polymerase Mr=450,000 E) serum albumin M,= M,= M, = 13,000 13,700 68,500

Explanation / Answer

To get the Kd value for HABA first react the maximum fractional saturation for HABA max value I read is 0.9, divide this value by 2 to get 0.45

Match this y value to x , the value of x is approximately 5 uM

Kd is the concentration complex ligand when fractional occupancy is 50%

Similarly we can stimate the Kd for the curve of 2-iminibiotin

It reaches a max fractional saturation of 1, divide this by 2 to get 0.5 and match this value to the concentration this is the value of kd , this value is approximately 0.5 a very small value

The higher the value of Kd the looser the binding

Ligands with small values of Kd bind tightly

So the 2IMB binds tightly

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