In Figure 4C, we see the inhibition of LRRK2 kinase activity by nanobody 1 (Nb1). Using the Lineweaver-Burke plot, estimate the Km of LRRK2 in the absence of Nb1. Show your work and provide your answer with the correct units.
In Figure 4C, we see the inhibition of LRRK2 kinase activity by nanobody 1 (Nb1). Using the Lineweaver-Burke plot, estimate the Km of LRRK2 in the absence of Nb1. Show your work and provide your answer with the correct units.
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
In Figure 4C, we see the inhibition of LRRK2 kinase activity by nanobody 1 (Nb1). Using the Lineweaver-Burke plot, estimate the Km of LRRK2 in the absence of Nb1. Show your work and provide your answer with the correct units.
![A
Arm
Ank LRR
Arm
Ank
LRR
2527
2527
Roc! COR
Kinase WD
Roc! COR
Kinase
WD
Rab P
1.54
: peptide substrate
1.5-
1.0-
1.0-
0.5-
0.5-
0.0
0.0
C
E
Nb1
Nb6
120-
Nb23
120-
宽1201
100-
100-
80-
80-
80-
60-
60-
60
40-
S 40-
40-
20-
|C50
20-
20-
IC 50= 8 ± 2 µM
IC 50= 14 ± 3 µuM
0-
65 μΜ
-2
-1
1
2
3
-3
-2
-1
1
2
-2
-1
1
2
3
Log [Nb1] (uM)
Log [Nb6] (uM)
Log [Nb23] (µM)
KaPP= 16 ± 4 µM
K
app= 5±1 µM
Kapp= 9 + 1 uM
40000-
100000-
a = 1.8 ± 0.5
a = 1.6 ± 0.4
a = 8 ± 2
40000-
30000-
75000-
30000-
50000-
20000-
20000 -
25000-
10000-
10000-
200
400
600
800
1000
200
400
600
800
1000
200
400
600
800
1000
[ATP] (uM)
[AΤΡΙ (μΜ)
[AΤPΙ (μΜ)
0.00010
0.0008
0.0006-
0.00005-
0.0004-
0.0003-
-0.02
-0.01
0.01
0.02
0.03
-0.02
0.01
0.01
0.02
0.03
-0.02
-0.01
0.01
0.02
0.03
[1/ATP] (µM1)
[1/ATP] (µM1)
[1/ATP] (µM^1)
-- (0 μΜ)
- (2.5 µM)
(5 μΜ)
(20 μΜ)
(50 μΜ)
(0 μΜ)
(2.5 μΜ )
(5 µM)
(20 μΜ)
(50 μΜ)
- (0 μΜ)
(10 μΜ)
( 30 μΜ)
(60 μΜ)
(100 μΜ)
Nb17
% Activity
Nb36
Nb38
Nb39
Nb42
relative velocity
Nb1
Nb6
Nb22
Nb23
Nb40
1/rel. velocity
MLi-2
No Nb
% Activity
relative velocity
Nb17
Nb36
Nb38
1/rel. velocity
Nb39
Nb42
Nb1
3
Nb6
% Activity
Nb22
Nb23
Nb40
relative velocity
MLi-2
No Nb'
1/rel. velocity](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2F17594ac0-a9a0-4ef8-a5f1-293992d00aab%2Fd6cf0976-6d0b-4f1c-b2c2-f699a1f6a817%2F97gpbqn_processed.jpeg&w=3840&q=75)
Transcribed Image Text:A
Arm
Ank LRR
Arm
Ank
LRR
2527
2527
Roc! COR
Kinase WD
Roc! COR
Kinase
WD
Rab P
1.54
: peptide substrate
1.5-
1.0-
1.0-
0.5-
0.5-
0.0
0.0
C
E
Nb1
Nb6
120-
Nb23
120-
宽1201
100-
100-
80-
80-
80-
60-
60-
60
40-
S 40-
40-
20-
|C50
20-
20-
IC 50= 8 ± 2 µM
IC 50= 14 ± 3 µuM
0-
65 μΜ
-2
-1
1
2
3
-3
-2
-1
1
2
-2
-1
1
2
3
Log [Nb1] (uM)
Log [Nb6] (uM)
Log [Nb23] (µM)
KaPP= 16 ± 4 µM
K
app= 5±1 µM
Kapp= 9 + 1 uM
40000-
100000-
a = 1.8 ± 0.5
a = 1.6 ± 0.4
a = 8 ± 2
40000-
30000-
75000-
30000-
50000-
20000-
20000 -
25000-
10000-
10000-
200
400
600
800
1000
200
400
600
800
1000
200
400
600
800
1000
[ATP] (uM)
[AΤΡΙ (μΜ)
[AΤPΙ (μΜ)
0.00010
0.0008
0.0006-
0.00005-
0.0004-
0.0003-
-0.02
-0.01
0.01
0.02
0.03
-0.02
0.01
0.01
0.02
0.03
-0.02
-0.01
0.01
0.02
0.03
[1/ATP] (µM1)
[1/ATP] (µM1)
[1/ATP] (µM^1)
-- (0 μΜ)
- (2.5 µM)
(5 μΜ)
(20 μΜ)
(50 μΜ)
(0 μΜ)
(2.5 μΜ )
(5 µM)
(20 μΜ)
(50 μΜ)
- (0 μΜ)
(10 μΜ)
( 30 μΜ)
(60 μΜ)
(100 μΜ)
Nb17
% Activity
Nb36
Nb38
Nb39
Nb42
relative velocity
Nb1
Nb6
Nb22
Nb23
Nb40
1/rel. velocity
MLi-2
No Nb
% Activity
relative velocity
Nb17
Nb36
Nb38
1/rel. velocity
Nb39
Nb42
Nb1
3
Nb6
% Activity
Nb22
Nb23
Nb40
relative velocity
MLi-2
No Nb'
1/rel. velocity
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