4. You have a protein that needs to bind ATP to be functional and has a...
4. You have a protein that needs to bind ATP to be functional and has a KD for ATP of 10 μM. In you experiment your protein concentration is much less than that of ATP, and your final ATP concentration is 30 μM.
A) What fraction of the protein is functional? Show your work.
B) Suppose that you have a fluorescent ATP analog that can bind and activate your proteins that becomes quenched when it binds. Why would fluorescence change upon binding?
C) Describe how you might use this fluorescence quenching to determine the Kd for ATP binding in the first place
D) Could fluorescence anisotropy work also? How would this experiment work?
Homework Answers
![0 = bending site uccupied . [PL] total binding sites [PL] & [P] [P]+(2] = [PL] we know, ka - Pil - [PL]= ka [P][2] Leo - ka (](http://img.homeworklib.com/questions/d2ec2670-e8c2-11eb-b10d-8b3d6f7e7ba5.png?x-oss-process=image/resize,w_560)
In
fluorescence quenching complex formation occurs between the
quencher molecule. Such complex cannot undergo excitation and the
fluorescence intensity decreases.
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