26 Jun I have so much trouble with this homework i dont know how to draw
Question
Enzyme kinetics homework
Total 75 points
Due at the beginning of class on 3/24/15
Name ________________________________
Signature _____________________________
Instructions: Answer all questions within this file (additional pages are acceptable if necessary). All answers
must be TYPED. You are free to work together, but please use your own words in answering the questions.
Turn in your own printed version of this file with answers embedded into the blank areas. All plots should be
done in Microsoft Excel and pasted into this document.
[SUBSTRATE] In mM
Assignment. A terrible plague of H3N2 influenza has begun to decimate the earths population. You convince
your boss at your pharmaceutical company to let you work on developing a new neuraminidase inhibitor to
combat this new strain. You make several hundred potential inhibitors and assay them with the neuraminidase
isolated from this new strain. Frustratingly, none of the compounds has the required Ki of less than 2 M to
effectively inhibit this enzyme. Finally, in desperation attempt, you decide to make one last compound. You
then assay this compound with the viral neuraminidase (which is used in the assay at 1 nM concentration).
After doing a complete study at various substrate and inhibitor concentrations, you arrive at the
following data. The numbers inside the box are the reaction rates measured in nM/s. This table is an embedded
MS excel file that can easily be copied into Microsoft Excel by cutting and pasting.
0.0083
0.0233
0.0383
0.0533
0.0683
0.0833
0.1665
0.4665
0.7664
1.0665
1.3665
1.6665
0
16.34
48.28
68.71
96.15
116.24
144.68
242.72
452.97
604.61
681.36
735.03
766.52
[INHIBITOR]
0.00125
15.82
43.65
64.54
83.39
104.72
119.88
190.09
297.30
345.70
367.65
384.22
393.83
in mM
0.0025
15.73
39.58
63.38
82.49
94.76
104.25
152.70
218.60
241.39
253.04
257.05
261.78
0.00374
15.64
38.11
59.00
70.91
83.61
94.93
128.23
173.65
184.77
191.35
196.05
198.05
0.005
15.55
39.57
55.57
67.45
77.59
81.74
111.62
141.35
149.75
154.62
157.22
159.00
**Numbers inside the box are rates with the units nM/s**
Question #1
Using the data given, determine the Vmax, kcat, and Km for the substrate with no
inhibitor present (Be sure to include units!). Attach a rate vs. [S] plot and a Lineweaver-Burke plot
below. Describe how you determined these parameters. Show equations. (30 points)
1
Question #2
Perform a complete inhibitor study (rate vs. [S] plots at each concentration of
inhibitor, your plots should each have 5 lines) using Excel and the data within the table. Attach the rate
vs. [S] and Lineweaver-Burke plots below and label each curve with the inhibitor concentration of that
curve (20 points).
Question #3
Answer the following question. What type of inhibitor is this compound? (5 points)
Question #4
Using a secondary plot like we described in class, determine the K i for this
compound. Attach the secondary plot below and describe how you determined K i from this plot (20
points).
Question #5 Did your compound save the world from this outbreak? Circle one of the following.
2
Y
N
3
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