Asp48 E(-1) D(-2) H (c) ( To identify the critical enzyme resi- due:substrate interaction and determinant of sub- strate specificity, steady-state kinetic studies have D(-4) been carried out by site-specific mutagenesis of the enzyme and by systematically changing amino acid residues in the 11-amino acid phosphotyrosine con- taining substrate that simulate residues 988 - 998 of the epidermal growth factor receptor (EGFR 988-998). The table below compares the kinetic parameters of the wild type enzyme with phosphotyrosine pep- tides in which amino acid residues are substituted. In the adjacent diagram the structural interactions between active site residues are illustrated for the first 6 residues of the substrate. Identify which sub- strate residue is most sensitive to substitution and justify your conclusion on the basis of the kinetic parameters. Is the decrease in Kcat/ KM due to loss of catalytic reactivity or due to suboptimal position- ing of the phospho-Tyr residue in the active site? A(-3) Tyr46 Ser216 Arg47 Asp181 Substrate DADEpYLIPQQG DADAP YLIPQQG DAAEpYLIPQQG AAAAP YLIPQQG kcat 5-1 44.6 ± 1.8 39.8 ± 0.32 35.3 ± 0.22 34.7 ± 0.25 PTP1B Km Y(0) L(+1) Gln262 Phe 182 kcat/Km им 3.9 ± 0.9 13.7 ± 0.46 6.6 ± 0.22 52.7 ± 0.7 10-7 × (s¯¹ M¯¹) 1.1 ± 0.25 0.29 ± 0.01 0.53 ± 0.02 0.066 ± 0.001
Asp48 E(-1) D(-2) H (c) ( To identify the critical enzyme resi- due:substrate interaction and determinant of sub- strate specificity, steady-state kinetic studies have D(-4) been carried out by site-specific mutagenesis of the enzyme and by systematically changing amino acid residues in the 11-amino acid phosphotyrosine con- taining substrate that simulate residues 988 - 998 of the epidermal growth factor receptor (EGFR 988-998). The table below compares the kinetic parameters of the wild type enzyme with phosphotyrosine pep- tides in which amino acid residues are substituted. In the adjacent diagram the structural interactions between active site residues are illustrated for the first 6 residues of the substrate. Identify which sub- strate residue is most sensitive to substitution and justify your conclusion on the basis of the kinetic parameters. Is the decrease in Kcat/ KM due to loss of catalytic reactivity or due to suboptimal position- ing of the phospho-Tyr residue in the active site? A(-3) Tyr46 Ser216 Arg47 Asp181 Substrate DADEpYLIPQQG DADAP YLIPQQG DAAEpYLIPQQG AAAAP YLIPQQG kcat 5-1 44.6 ± 1.8 39.8 ± 0.32 35.3 ± 0.22 34.7 ± 0.25 PTP1B Km Y(0) L(+1) Gln262 Phe 182 kcat/Km им 3.9 ± 0.9 13.7 ± 0.46 6.6 ± 0.22 52.7 ± 0.7 10-7 × (s¯¹ M¯¹) 1.1 ± 0.25 0.29 ± 0.01 0.53 ± 0.02 0.066 ± 0.001
Human Heredity: Principles and Issues (MindTap Course List)
11th Edition
ISBN:9781305251052
Author:Michael Cummings
Publisher:Michael Cummings
Chapter10: From Proteins To Phenotypes
Section: Chapter Questions
Problem 9QP: a. Compounds A, B, C, and D are known to be intermediates in the pathway for production of protein...
Related questions
Question
![(c) (
due:substrate interaction and determinant of sub-
To identify the critical enzyme resi-
D(-2)
Arg47
strate specificity, steady-state kinetic studies have D(-4).
been carried out by site-specific mutagenesis of the
enzyme and by systematically changing amino acid
residues in the 11-amino acid phosphotyrosine con-
taining substrate that simulate residues 988 – 998
of the epidermal growth factor receptor
(EGFR988–998).
Asp48
E(-1)
A(-3)
The table below compares the kinetic parameters
of the wild type enzyme with phosphotyrosine pep-
tides in which amino acid residues are substituted.
In the adjacent diagram the structural interactions
L(+1)
Tyr46
Gln262
Y(0)
between active site residues are illustrated for the
first 6 residues of the substrate. Identify which sub-
strate residue is most sensitive to substitution and
Ser216
justify your conclusion on the basis of the kinetic
parameters. Is the decrease in kcat/ KM due to loss
of catalytic reactivity or due to suboptimal position-
ing of the phospho-Tyr residue in the active site?
Phe182
Asp181
PTP1B
Substrate
kcat
K.
kcat/Km
JUM
10-7 x (s-1 M1)
DADEPYLIPQQG
DADAPYLIPQQG
DAAEPYLIPQQG
AAAAPYLIPQQG
44.6 ± 1.8
39.8 + 0.32
3.9 + 0.9
13.7 + 0.46
1.1 + 0.25
0.29 + 0.01
35.3 + 0.22
6.6 ± 0.22
0.53 + 0.02
34.7 + 0.25
52.7 + 0.7
0.066 0.001](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2F7e3218aa-1b65-42a2-9071-7edb43c3a555%2F02fe31f2-2457-45c7-a788-5fed0e9d2ea2%2Fckpg7ef_processed.png&w=3840&q=75)
Transcribed Image Text:(c) (
due:substrate interaction and determinant of sub-
To identify the critical enzyme resi-
D(-2)
Arg47
strate specificity, steady-state kinetic studies have D(-4).
been carried out by site-specific mutagenesis of the
enzyme and by systematically changing amino acid
residues in the 11-amino acid phosphotyrosine con-
taining substrate that simulate residues 988 – 998
of the epidermal growth factor receptor
(EGFR988–998).
Asp48
E(-1)
A(-3)
The table below compares the kinetic parameters
of the wild type enzyme with phosphotyrosine pep-
tides in which amino acid residues are substituted.
In the adjacent diagram the structural interactions
L(+1)
Tyr46
Gln262
Y(0)
between active site residues are illustrated for the
first 6 residues of the substrate. Identify which sub-
strate residue is most sensitive to substitution and
Ser216
justify your conclusion on the basis of the kinetic
parameters. Is the decrease in kcat/ KM due to loss
of catalytic reactivity or due to suboptimal position-
ing of the phospho-Tyr residue in the active site?
Phe182
Asp181
PTP1B
Substrate
kcat
K.
kcat/Km
JUM
10-7 x (s-1 M1)
DADEPYLIPQQG
DADAPYLIPQQG
DAAEPYLIPQQG
AAAAPYLIPQQG
44.6 ± 1.8
39.8 + 0.32
3.9 + 0.9
13.7 + 0.46
1.1 + 0.25
0.29 + 0.01
35.3 + 0.22
6.6 ± 0.22
0.53 + 0.02
34.7 + 0.25
52.7 + 0.7
0.066 0.001
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