16 Jun MCB 2000 Spring 2016 Problem Set #2-You measure the initial rate of an enzyme reaction
Question
MCB 2000
Spring 2016
Problem Set #2
MCB 2000 Problem Set #2
(Note: You must show your work to receive full credit!)
Due Date: Week of March 7th at the beginning of your lab section
Enzyme kinetic problems
1. You measure the initial rate of an enzyme reaction as a function of substrate concentration in the
presence and absence of an inhibitor. The following data was obtained:
[S]
(mM)
Vo (-) Inhibitor
(mM/min)
Vo (+) Inhibitor
(mM/min)
0.0001
0.0002
0.0005
0.001
0.002
33
50
71
83
91
17
29
50
67
80
Please submit calculations and graph for full credit!
Note: You are required to use Excel to generate the Lineweaver-Burk plot
(a) (10 points) Create a Lineweaver-Burk plot (inhibited and uninhibited should be on the
same plot!)
(b) (15 points) Calculate the Vmax in the presence and absence of the inhibitor. Show your
calculation.
(c) (15 points) Calculate the Km in the presence and absence of the inhibitor. Show your
calculation
(d) (10 points) What type of inhibitor is this? How do you know (clearly explain)?
(ADDITIONAL QUESTIONS ON BACK!!!!)
MCB 2000
Spring 2016
Problem Set #2
2. Prostaglandins are derivatives of fatty acids that are responsible for producing fever, inflammation
and its associated pain. They are synthesized from a 20 carbon fatty acid, arachidonic acid (AA), in a
reaction catalyzed by prostaglandin endoperoxide synthase. The product of the reaction, PGG 2, can
then be used to synthesize other prostaglandins. Ibuprofen is an inhibitor of prostaglandin
endoperoxide synthase, thus it used to relieve pain and inflammation.
The following data was generated in the laboratory:
Arachidonic Acid
(mM)
Rate of formation of PGG2
(mM/min)
Rate of formation of PGG2 with
10 mg/ml ibuprofen
(mM/min)
0.5
1.0
1.5
2.5
3.5
23.5
32.2
36.9
41.8
44.0
16.67
25.25
30.49
37.04
38.91
Please submit calculations and graph for full credit!
Note: You are required to use Excel to generate the Lineweaver-Burk plot
(a) (15 points) Create a Lineweaver-Burk plot (inhibited and uninhibited should be on the
same plot!)
(b) (15 points)Calculate Vmax and Km in absence of the inhibitor. Show your calculations.
(c) (20 points) Calculate the Vmax and Km in the presence of the inhibitor. Show your
calculations. What type of inhibitor is ibuprofen? Briefly explain your answer.
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