You start by conducting a binding experiment using purified Slugase enzyme at a concentration of 100 nM. You add varying amounts of radioactively labeled tannin ligand and mix the reactions with a resin that binds to proteins (including Slugase), which can be pelleted in a centrifuge. The amount of radioactivity in the pellet represents the amount of tannin-Slugase complex. You also perform a control experiment in which you do not add any Slugase to the mixture and find that you get some signal in the pellet that is non- specific. Using this approach, you generate the following data: [tannin], μM 0.1 0.5 1.0 2.0 4.0 8.0 16.0 32.0 64.0 Observed Binding, radioactive counts 4.22 19.00 33.83 55.55 82.00 108.35 131.08 151.21 174.22 Non-specific Binding, radioactive counts 0.05 0.25 0.50 1.00 2.00 4.00 8.00 16.00 32.00 Specific Fractional Binding, Saturation radioactive counts 4.17 18.75 33.33 54.55 80.00 104.35 123.08 135.21 142.22 (a) In the space below, make a plot of the [tannin] on the x-axis and the appropriate data for Slugase binding on the y-axis. Using this plot, estimate what the value of the dissociation constant (Kd) is for the Slugase-tannin interaction.
You start by conducting a binding experiment using purified Slugase enzyme at a concentration of 100 nM. You add varying amounts of radioactively labeled tannin ligand and mix the reactions with a resin that binds to proteins (including Slugase), which can be pelleted in a centrifuge. The amount of radioactivity in the pellet represents the amount of tannin-Slugase complex. You also perform a control experiment in which you do not add any Slugase to the mixture and find that you get some signal in the pellet that is non- specific. Using this approach, you generate the following data: [tannin], μM 0.1 0.5 1.0 2.0 4.0 8.0 16.0 32.0 64.0 Observed Binding, radioactive counts 4.22 19.00 33.83 55.55 82.00 108.35 131.08 151.21 174.22 Non-specific Binding, radioactive counts 0.05 0.25 0.50 1.00 2.00 4.00 8.00 16.00 32.00 Specific Fractional Binding, Saturation radioactive counts 4.17 18.75 33.33 54.55 80.00 104.35 123.08 135.21 142.22 (a) In the space below, make a plot of the [tannin] on the x-axis and the appropriate data for Slugase binding on the y-axis. Using this plot, estimate what the value of the dissociation constant (Kd) is for the Slugase-tannin interaction.
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
Related questions
Question
Solve part a
![radioactive counts
200
150
100
50
0
0
10
20
30
[Tannin], uM
40
50
60](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2Fd6957468-6fb1-4659-9083-987b06676d6d%2F81d1eac1-cdf8-41da-884c-167746fa9c75%2Fz61pkil_processed.jpeg&w=3840&q=75)
Transcribed Image Text:radioactive counts
200
150
100
50
0
0
10
20
30
[Tannin], uM
40
50
60
![You start by conducting a binding experiment using purified Slugase enzyme at a
concentration of 100 nM. You add varying amounts of radioactively labeled tannin ligand
and mix the reactions with a resin that binds to proteins (including Slugase), which can be
pelleted in a centrifuge. The amount of radioactivity in the pellet represents the amount of
tannin-Slugase complex. You also perform a control experiment in which you do not add
any Slugase to the mixture and find that you get some signal in the pellet that is non-
specific.
Using this approach, you generate the following data:
[tannin], µM
0.1
0.5
1.0
2.0
4.0
8.0
16.0
32.0
64.0
Observed
Binding,
radioactive
counts
4.22
19.00
33.83
55.55
82.00
108.35
131.08
151.21
174.22
Non-specific
Binding,
radioactive
counts
0.05
0.25
0.50
1.00
2.00
4.00
8.00
16.00
32.00
Specific Fractional
Binding, Saturation
radioactive
counts
4.17
18.75
33.33
54.55
80.00
104.35
123.08
135.21
142.22
(a) In the space below, make a plot of the [tannin] on the x-axis and the appropriate data for
Slugase binding on the y-axis. Using this plot, estimate what the value of the
dissociation constant (Kd) is for the Slugase-tannin interaction.](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2Fd6957468-6fb1-4659-9083-987b06676d6d%2F81d1eac1-cdf8-41da-884c-167746fa9c75%2F0f8ejbq_processed.jpeg&w=3840&q=75)
Transcribed Image Text:You start by conducting a binding experiment using purified Slugase enzyme at a
concentration of 100 nM. You add varying amounts of radioactively labeled tannin ligand
and mix the reactions with a resin that binds to proteins (including Slugase), which can be
pelleted in a centrifuge. The amount of radioactivity in the pellet represents the amount of
tannin-Slugase complex. You also perform a control experiment in which you do not add
any Slugase to the mixture and find that you get some signal in the pellet that is non-
specific.
Using this approach, you generate the following data:
[tannin], µM
0.1
0.5
1.0
2.0
4.0
8.0
16.0
32.0
64.0
Observed
Binding,
radioactive
counts
4.22
19.00
33.83
55.55
82.00
108.35
131.08
151.21
174.22
Non-specific
Binding,
radioactive
counts
0.05
0.25
0.50
1.00
2.00
4.00
8.00
16.00
32.00
Specific Fractional
Binding, Saturation
radioactive
counts
4.17
18.75
33.33
54.55
80.00
104.35
123.08
135.21
142.22
(a) In the space below, make a plot of the [tannin] on the x-axis and the appropriate data for
Slugase binding on the y-axis. Using this plot, estimate what the value of the
dissociation constant (Kd) is for the Slugase-tannin interaction.
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