Any feedback?
Information on EC 6.1.1.1 - tyrosine-tRNA ligase and Organism(s) Escherichia coli and UniProt Accession P0AGJ9
for references in articles please use BRENDA:EC6.1.1.1Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
6.1.1.1 tyrosine-tRNA ligaseSpecify your search results
Word Map
- 6.1.1.1
- synthetases
- aminoacylation
-
tyrosylation
- stearothermophilus
-
anticodon
-
amber
- jannaschii
- trprs
-
aarss
- methanococcus
-
kmsks
-
tryptophanyl-trna
-
charcot-marie-tooth
-
mischarging
-
trna-like
- d-tyrosine
-
self-splicing
- methanocaldococcus
-
half-of-the-sites
- elr
-
non-cognate
- methionyl-trna
-
isoleucyl-trna
- medicine
- biotechnology
-
sideroblastic
-
anticodon-binding
-
tyrts
- 3-iodo-l-tyrosine
- glycyl-trna
- drug development
- pharmacology
- leurs
-
misacylation
- phenylalanyl-trna
-
fersht
-
brome
- hisrs
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
tyrosyl-trna synthetase, tyrrs, cyt-18, mitochondrial tyrosyl-trna synthetase, mini-tyrrs, tyrrss, cyt-18 protein, tyrosyl trna synthetase, mttyrrs, ldtyrrs, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
Tyrosine translase
-
-
-
-
Tyrosine tRNA synthetase
Tyrosine--tRNA ligase
-
-
-
-
Tyrosine-transfer ribonucleate synthetase
-
-
-
-
Tyrosine-transfer RNA ligase
-
-
-
-
Tyrosyl--tRNA ligase
-
-
-
-
Tyrosyl-transfer ribonucleate synthetase
-
-
-
-
Tyrosyl-transfer ribonucleic acid synthetase
-
-
-
-
Tyrosyl-transfer RNA synthetase
-
-
-
-
Tyrosyl-tRNA ligase
-
-
-
-
Tyrosyl-tRNA synthetase
TyrRS
Tyrosine tRNA synthetase
-
-
-
-
Tyrosyl-tRNA synthetase
-
-
-
-
TyrRS
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + L-tyrosine + tRNATyr = AMP + diphosphate + L-tyrosyl-tRNATyr
mechanism
-
ATP + L-tyrosine + tRNATyr = AMP + diphosphate + L-tyrosyl-tRNATyr
amino acid residues Y37 and Q195 are involved in substrate specificity determination
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
esterification
Acylation
esterification
-
-
-
-
Acylation
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
L-tyrosine:tRNATyr ligase (AMP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY
9023-45-4
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + 3-azido-L-tyrosine + tRNATyr
AMP + diphosphate + 3-azido-L-tyrosyl-tRNATyr
-
-
-
?
ATP + 3-iodo-L-tyrosine + tRNATyr
AMP + diphosphate + 3-iodo-L-tyrosyl-tRNATyr
-
-
-
?
ATP + D-tyrosine + tRNATyr
AMP + diphosphate + D-tyrosyl-tRNATyr
enzyme TyrRS has a detectable, natural activity for the D-tyrosine stereoisomer, only tenfold less than for L-Tyr
-
-
?
ATP + 3-iodo-L-tyrosine + tRNATyr
AMP + L-Tyr-tRNATyr + diphosphate
-
mutant Y73V/Q195C and other mutants, no activity with the wild-type enzyme
-
?
ATP + L-beta-(5-hydroxy-2-pyridyl)-alanine + tRNATyr
AMP + L-beta-(5-hydroxy-2-pyridyl)-alanine-tRNATyr + diphosphate
ATP + L-tyrosine + tRNATyr
AMP + diphosphate + L-tyrosyl-tRNATyr
-
-
-
?
ATP + L-tyrosine + tRNATyr
AMP + diphosphate + L-tyrosyl-tRNATyr
-
-
-
?
ATP + L-beta-(5-hydroxy-2-pyridyl)-alanine + tRNATyr
AMP + L-beta-(5-hydroxy-2-pyridyl)-alanine-tRNATyr + diphosphate
-
L-beta-(5-hydroxy-2-pyridyl)-alanine i.e. azatyrosine, mutant F130S, and wild-type enzyme the latter showing low activity
-
?
ATP + L-beta-(5-hydroxy-2-pyridyl)-alanine + tRNATyr
AMP + L-beta-(5-hydroxy-2-pyridyl)-alanine-tRNATyr + diphosphate
-
L-beta-(5-hydroxy-2-pyridyl)-alanine i.e. azatyrosine, mutant F130S shows 17fold higher activity in vivo than the wild-type enzyme
-
?
ATP + tyrosine + tRNATyr
AMP + Tyr-tRNATyr + diphosphate
-
same affinity for tRNA or tRNA acylated with tyrosine
-
-
?
additional information
?
-
residues Asp81, Tyr175, Gln179, and Gln201 coordinate the ammonium group of the L-Tyr ligand
-
-
?
additional information
?
-
-
residues Asp81, Tyr175, Gln179, and Gln201 coordinate the ammonium group of the L-Tyr ligand
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + L-beta-(5-hydroxy-2-pyridyl)-alanine + tRNATyr
AMP + L-beta-(5-hydroxy-2-pyridyl)-alanine-tRNATyr + diphosphate
-
L-beta-(5-hydroxy-2-pyridyl)-alanine i.e. azatyrosine, mutant F130S shows 17fold higher activity in vivo than the wild-type enzyme
-
?
ATP + L-tyrosine + tRNATyr
AMP + diphosphate + L-tyrosyl-tRNATyr
-
-
-
?
ATP + L-tyrosine + tRNATyr
AMP + diphosphate + L-tyrosyl-tRNATyr
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Mg2+
Mg2+
-
required, spermidine can substitute for Mg2+ in the transacylation reaction, i.e. transfer of tyrosine from the tyrosine-AMP-enzyme complex to tRNA, but not in the tyrosine activation reaction, i.e. formation of the tyrosine-AMP-enzyme complex from tyrosine + ATP + enzyme
Mg2+
-
required, shows regulatory function, binding of 1 Mg2+ per tRNA molecule for the transfer reaction, binding to the tRNA is weakened by chloride
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
chloride
-
inhibition in presence of 1 mM free Mg2+, no inhibition in presence of 10 mM free Mg2+
additional information
natural compounds as inhibitors of tyrosyl-tRNA synthetase, effects of various polyphenols, alkaloids, and terpenes-secondary metabolites produced by higher plants, overview. Most of them act as competitive inhibitors. Structure-activity relationship shows that the most potent flavonoid inhibitors contain hydroxyl group at position 5 and 7 of A ring and a -OCH3 group at position 4' of B ring
-
additional information
-
natural compounds as inhibitors of tyrosyl-tRNA synthetase, effects of various polyphenols, alkaloids, and terpenes-secondary metabolites produced by higher plants, overview. Most of them act as competitive inhibitors. Structure-activity relationship shows that the most potent flavonoid inhibitors contain hydroxyl group at position 5 and 7 of A ring and a -OCH3 group at position 4' of B ring
-
additional information
-
no inhibition by acetate up to 200 mM in presence of 1 mM free Mg2+
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00043
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant wild-type enzyme
0.00045
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 144AcK
0.00049
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 355AcK
0.00071
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 235AcK
0.00223
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 85AcK
0.00669
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 355AcK
0.00678
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 85AcK
0.00713
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant wild-type enzyme
0.00728
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 144AcK
0.00845
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 235AcK
0.00037
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant wild-type enzyme
0.00042
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant lysine acetylated enzyme mutant 144AcK
0.00045
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant lysine acetylated enzyme mutant 85AcK
0.00048
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant lysine acetylated enzyme mutant 355AcK
additional information
additional information
calculation of the L-Tyr/D-Tyr binding free energy difference. Michaelis-Menten kinetics of wild-type and mutant enzymes
-
additional information
additional information
-
calculation of the L-Tyr/D-Tyr binding free energy difference. Michaelis-Menten kinetics of wild-type and mutant enzymes
-
additional information
additional information
-
stoichiometry of substrate binding
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.1
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 235AcK
2
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 85AcK
11.9
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 355AcK
12.3
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 144AcK
14.2
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant wild-type enzyme
0.1
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 235AcK
2
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 85AcK
11.9
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 355AcK
12.3
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 144AcK
14.2
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant wild-type enzyme
0.15
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant lysine acetylated enzyme mutant 85AcK
1.09
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant lysine acetylated enzyme mutant 355AcK
1.17
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant lysine acetylated enzyme mutant 144AcK
1.32
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant wild-type enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
140
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 235AcK
900
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 85AcK
24290
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 355AcK
27330
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 144AcK
33020
ATP
pH 7.2, 37°C, Tyr activation activity, recombinant wild-type enzyme
10
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 235AcK
290
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 85AcK
1570
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 144AcK
1780
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant lysine acetylated enzyme mutant 355AcK
1990
L-tyrosine
pH 7.2, 37°C, Tyr activation activity, recombinant wild-type enzyme
0.33
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant lysine acetylated enzyme mutant 85AcK
2.27
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant lysine acetylated enzyme mutant 355AcK
2.78
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant lysine acetylated enzyme mutant 144AcK
3.51
tRNATyr
pH 7.2, 37°C, tRNATyr aminoacylation activity, recombinant wild-type enzyme
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
additional information
malfunction
lysine acetylation can be a possible mechanism for modulating aminoacyl-tRNA synthetases enzyme activities, thus affecting translation. Of recombinantly expressed site-specifically acetylated TyrRS variants, TyrRS-85AcK and -235AcK show dramatic decreases in activity. Variant TyrRS-238AcK has no detectable activity, while variants TyrRS-144AcK and -355AcK have similar activities compared to the wild-type TyrRS. TyrRS-85AcK has a fivefold increase in the KM value for ATP, indicating its role in ATP binding. TyrRS-235AcK has slightly changed KM values for both ATP and tyrosine but a 200fold decrease in catalytic efficiency, suggesting its role in catalysis. K235 and K238 of TyrRS characterized in this study are the two lysine residues in the KMSKS motif. Kinetics for acetylated mutant variants, overview
malfunction
of six mutants tested, two are active towards D-Tyr; one of these has an inverted stereospecificity, with a large preference for D-Tyr, but its activity is low
additional information
lysine acetylation could be a possible mechanism for modulating aminoacyl-tRNA synthetases enzyme activities, thus affecting translation
additional information
-
lysine acetylation could be a possible mechanism for modulating aminoacyl-tRNA synthetases enzyme activities, thus affecting translation
additional information
the TyrRS stereospecificity is robust towards charge rearrangements near the ligand. Whereas most aminoacyl-tRNA synthetases (aaRSs) have a strong preference for their L-amino acid substrate, TyrRS has a detectable, natural activity for the D-tyrosine stereoisomer, only tenfold less than for L-Tyr; additional protection against D-Tyr is usually provided by another enzyme, D-aminoacyl-tRNA hydrolase. Enzyme molecular dynamics simulations using the crystal structure of Escherichia coli TyrRS bound to a tyrosyl adenylate analogue, PDB ID 1VBM
additional information
-
the TyrRS stereospecificity is robust towards charge rearrangements near the ligand. Whereas most aminoacyl-tRNA synthetases (aaRSs) have a strong preference for their L-amino acid substrate, TyrRS has a detectable, natural activity for the D-tyrosine stereoisomer, only tenfold less than for L-Tyr; additional protection against D-Tyr is usually provided by another enzyme, D-aminoacyl-tRNA hydrolase. Enzyme molecular dynamics simulations using the crystal structure of Escherichia coli TyrRS bound to a tyrosyl adenylate analogue, PDB ID 1VBM
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
three-dimensional modeling of the Escherichia coli enzyme constructed on the basis of the X-ray crystal structure of Bacillus stearothermophilus enzyme complexed with tyrosinyl adenylate, PDB code 3TS1
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
acetylation
tyrosyl-tRNA synthetase (TyrRS) in Escherichia coli is acetylated at multiple lysine residues. Acetylation at K85, K235, and K238 impairs the enzyme activity, by genetic-code-expansion strategy to site-specifically incorporate Nepsilonacetyl-l-lysine into selected positions of TyrRS for in vitro characterization. Lysine residue K355 is never acetylated. Most acetylated lysine residues in TyrRS are sensitive to the Escherichia coli deacetylase CobB but not YcgC, overview. Mapping of acetylated lysine residues on the crystal structure of TyrRS, PDB ID 1VBM
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor-diffusion method. Crystal structures of TyrRS catalytic domain, in complex with L-tyrosine and L-tyrosyladenylate analogue, 5'-O-[N-(L-tyrosyl)sulfamoyl]adenosine, are solved at 2.0 A and 2.7 A resolution
the iodoTyrRS-ec-3-azide-L-tyrosine structure is determined at a resolution of 1.8 A
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D81H/Q179E/Q201D
site-directed mutagenesis, the mutant shows no activity for L-Tyr and D-Tyr
D81K/Q179E
site-directed mutagenesis, the mutant shows no activity for L-Tyr and D-Tyr
D81K/Q179E/Q201D
site-directed mutagenesis, the mutant shows no activity for L-Tyr and D-Tyr
D81N
site-directed mutagenesis, the mutant shows a preference for L-Tyr that is much stronger than for the wild-type TyrRS. The ligand ammonium is coordinated at the L-Tyr endpoint by Asp41 (which replaces Asp81 in the coordination shell) and Tyr175 but not Gln179. At the D-Tyr endpoint, the ligand ammonium is coordinated by a mixture of Gln201, Tyr175, Asp41, and sometimes weakly by Gln179
D81R
site-drected mutagenesis, the mutant shows low activity for L-Tyr and D-Tyr, and the same KM value for L-Tyr compared to wild-type enzyme, the mutant is D-Tyr specific, but with low activity
F130S
-
construction of a plasmid library of randomly mutated gene tyrS by PCR, isolation of a mutant R-6-A-7 which incorporates L-beta-(5-hydroxy-2-pyridyl)-alanine in transformed Escherichia coli cells in vivo, increased temperature instability
Q179A
-
site-directed mutagenesis, reduced activity with L-tyrosine, no activity with 3-iodo-L-tyrosine
Q179N
-
site-directed mutagenesis, reduced activity with L-tyrosine, no activity with 3-iodo-L-tyrosine
Q179S
-
site-directed mutagenesis, reduced activity with L-tyrosine, no activity with 3-iodo-L-tyrosine
Q195A
-
site-directed mutagenesis, active with L-tyrosine, low activity with 3-iodo-L-tyrosine
Q195C
-
site-directed mutagenesis, reduced activity with L-tyrosine, low activity with 3-iodo-L-tyrosine
Q195D
-
site-directed mutagenesis, highly reduced activity with L-tyrosine, low activity with 3-iodo-L-tyrosine
Q195E
-
site-directed mutagenesis, active with L-tyrosine, no activity with 3-iodo-L-tyrosine
Q195G
-
site-directed mutagenesis, reduced activity with L-tyrosine, no activity with 3-iodo-L-tyrosine
Q195H
-
site-directed mutagenesis, active with L-tyrosine, no activity with 3-iodo-L-tyrosine
Q195I
-
site-directed mutagenesis, highly reduced activity with L-tyrosine, no activity with 3-iodo-L-tyrosine
Q195L
-
site-directed mutagenesis, reduced activity with L-tyrosine, no activity with 3-iodo-L-tyrosine
Q195M
-
site-directed mutagenesis, highly reduced activity with L-tyrosine, no activity with 3-iodo-L-tyrosine
Q195N
-
site-directed mutagenesis, reduced activity with L-tyrosine, low activity with 3-iodo-L-tyrosine
Q195S
-
site-directed mutagenesis, active with L-tyrosine, low activity with 3-iodo-L-tyrosine
Q195T
-
site-directed mutagenesis, active with L-tyrosine, no activity with 3-iodo-L-tyrosine
Q195V
-
site-directed mutagenesis, reduced activity with L-tyrosine, no activity with 3-iodo-L-tyrosine
Q195Y
-
site-directed mutagenesis, reduced activity with L-tyrosine, no activity with 3-iodo-L-tyrosine
Y37A/Q195C
-
site-directed mutagenesis, active with L-tyrosine and 3-iodo-L-tyrosine, preference for the latter, reduced overall activity
Y37A/Q195S
-
site-directed mutagenesis, equally low activity with L-tyrosine and 3-iodo-L-tyrosine
Y37I/Q195A
-
site-directed mutagenesis, equally low activity with L-tyrosine and 3-iodo-L-tyrosine
Y37L/Q195A
-
site-directed mutagenesis, highly reduced activity with L-tyrosine, no activity with 3-iodo-L-tyrosine
Y37L/Q195C
-
site-directed mutagenesis, equally low activity with L-tyrosine and 3-iodo-L-tyrosine
Y37L/Q195N
-
site-directed mutagenesis, highly reduced activity with L-tyrosine, no activity with 3-iodo-L-tyrosine
Y37V/Q195N
-
site-directed mutagenesis, active with L-tyrosine and 3-iodo-L-tyrosine, preference for the latter, reduced overall activity
Y37V/Q195S
-
site-directed mutagenesis, reduced activity with L-tyrosine and 3-iodo-L-tyrosine
Y73A
-
site-directed mutagenesis, equally active with L-tyrosine and 3-iodo-L-tyrosine
Y73F
-
site-directed mutagenesis, active with L-tyrosine, no activity with 3-iodo-L-tyrosine
Y73G
-
site-directed mutagenesis, equally active with L-tyrosine and 3-iodo-L-tyrosine, reduced overall activity
Y73H
-
site-directed mutagenesis, active with L-tyrosine, and slightly active with 3-iodo-L-tyrosine
Y73I
-
site-directed mutagenesis, active with L-tyrosine and 3-iodo-L-tyrosine, preference for the latter
Y73L
-
site-directed mutagenesis, equally active with L-tyrosine and 3-iodo-L-tyrosine
Y73M
-
site-directed mutagenesis, equally active with L-tyrosine and 3-iodo-L-tyrosine
Y73S
-
site-directed mutagenesis, active with L-tyrosine and 3-iodo-L-tyrosine, preference for the first
Y73V
-
site-directed mutagenesis, equally active with L-tyrosine and 3-iodo-L-tyrosine
Y73V/Q195A
-
site-directed mutagenesis, active with L-tyrosine and 3-iodo-L-tyrosine, preference for the latter, reduced overall activity
Y73V/Q195C
-
site-directed mutagenesis, 10fold more active with 3-iodo-L-tyrosine than with L-tyrosine, reduced overall activity
additional information
additional information
computational mutant design using the crystal structure of Escherichia coli TyrRS bound to a tyrosyl adenylate analogue, PDB ID 1VBM
additional information
-
computational mutant design using the crystal structure of Escherichia coli TyrRS bound to a tyrosyl adenylate analogue, PDB ID 1VBM
additional information
of recombinantly expressed site-specifically acetylated TyrRS variants, TyrRS-85AcK and -235AcK show dramatic decreases in activity. Variant TyrRS-238AcK has no detectable activity, while variants TyrRS-144AcK and -355AcK have similar activities compared to the wild-type TyrRS. TyrRS-85AcK has a fivefold increase in the KM value for ATP, indicating its role in ATP binding. TyrRS-235AcK has slightly changed KM values for both ATP and tyrosine but a 200fold decrease in catalytic efficiency, suggesting its role in catalysis
additional information
-
of recombinantly expressed site-specifically acetylated TyrRS variants, TyrRS-85AcK and -235AcK show dramatic decreases in activity. Variant TyrRS-238AcK has no detectable activity, while variants TyrRS-144AcK and -355AcK have similar activities compared to the wild-type TyrRS. TyrRS-85AcK has a fivefold increase in the KM value for ATP, indicating its role in ATP binding. TyrRS-235AcK has slightly changed KM values for both ATP and tyrosine but a 200fold decrease in catalytic efficiency, suggesting its role in catalysis
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
46
-
40% loss of activity in 4-12 min, depending on the bacterial strains used for purification, stabilization by substrates
50
-
10 min, wild-type enzyme is not affected, while the mutant F130S is almost completely inactivated
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 0.025 M potassium phosphate buffer, pH 6.7, 0.01 M 2-mercaptoethanol, 50% glycerol, no loss of activity in 6 months
-
4°C, 20 mM Tris-HCl buffer, pH 7, 80% loss of activity within 48 h, 5% loss of activity in presence of 10% glycerol and 3 mM 2-mercaptoethanol
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography to homogeneity, recombinant His-tagged mutant enzymes from Saccharomyces cerevisiae strain BY4742 by nickel affinity chromatography to over 90% purity
recombinant His6-tagged wild-type enzyme and site-specifically acetylated TyrRS variants from Escherichia coli strain BL21(DE3) cells
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene tyrS, recombinant expression of His-tagged wild-type enzyme in Escherichia coli strain BL21(DE3), recombinant expression of His-tagged mutant enzymes in Saccharomyces cerevisiae strain BY4742
gene tyrS, recombinant expression of His6-tagged wild-type enzyme and site-specifically acetylated TyrRS variants in Escherichia coli strain BL21(DE3) cells
gene tyrS, expression of wild-type and mutant enzymes as His-tagged proteins in strain JM109
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
lysine acetylation can be a possible mechanism for modulating aminoacyl-tRNA synthetases enzyme activities, thus affecting translation
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
the present engineering allows Escherichia coli TyrRS variants for non-natural amino acids to be developed in Escherichia coli, for use in both eukaryotic and bacterial cells for genetic code expansion
drug development
TyrRS enzymes are candidates for therapeutic targets in the prevention and therapy of microbial infections
biotechnology
-
site-specific incorporation of 3-iodo-L-tyrosine into proteins in a cell-free system for use in specialized in vitro translation systems
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Grosse, F.; Krauss, G.; Kownatzki, R.; Maass, G.
The binding of tyrosinyl-5'-AMP to tyrosyl synthetase (E. coli)
Nucleic Acids Res.
6
1631-1638
1979
Escherichia coli
Chousterman, S.; Chapeville, F.
Tyrosyl-tRNA synthetase of Escherichia coli B. Role of magnesium in the reaction catalyzed by the enzyme
Eur. J. Biochem.
35
46-50
1988
Escherichia coli
Chousterman, S.; Chapeville, F.
Escherichia coli tyrosyl-tRNA synthetase. Influence of magnesium ions on the enzme activity
FEBS Lett.
17
153-157
1971
Escherichia coli
Buonocore, V.; Harris, M.H.; Schlesinger, S.
Properties of tyrosyl transfer ribonucleic acid synthetase from two tyrS mutants of Escherichia coli K-12
J. Biol. Chem.
247
843-4849
1972
Escherichia coli
-
Chousterman, S.; Chapeville, F.
Tyrosyl-tRNA synthetase of Escherichia coli B
Eur. J. Biochem.
35
51-56
1973
Escherichia coli
Jakes, R.; Fersht, A.R.
Tyrosyl-tRNA synthetase from Escherichia coli. Stoichiometry of ligand binding and of half-of-the-sites reactivity in aminoacylation
Biochemistry
14
3344-3350
1975
Escherichia coli
Bruton, C.; Jakes, R.; Atkinson, T.
Gram-scale purification of methionyl-tRNA and tyrosyl-tRNA synthetases from Escherichia coli
Eur. J. Biochem.
59
327-333
1975
Escherichia coli
Airas, R.K.
Chloride affects the interaction between tyrosyl-tRNA synthetase and tRNA
Biochim. Biophys. Acta
1472
51-61
1999
Geobacillus stearothermophilus, Escherichia coli
Hamano-Takaku, F.; Iwama, T.; Saito-Yano, S.; Takaku, K.; Monden, Y.; Kitabatake, M.; Soll, D.; Nishimura, S.
A mutant Escherichia coli tyrosyl-tRNA synthetase utilizes the unnatural amino acid azatyrosine more efficiently than tyrosine
J. Biol. Chem.
275
40324-40328
2000
Escherichia coli
Kiga, D.; Sakamoto, K.; Kodama, K.; Kigawa, T.; Matsuda, T.; Yabuki, T.; Shirouzu, M.; Harada, Y.; Nakayama, H.; Takio, K.; Hasegawa, Y.; Endo, Y.; Hirao, I.; Yokoyama, S.
An engineered Escherichia coli tyrosyl-tRNA synthetase for site-specific incorporation of an unnatural amino acid into proteins in eukaryotic translation and its application in a wheat germ cell-free system
Proc. Natl. Acad. Sci. USA
99
9715-9720
2002
Escherichia coli
Kobayashi, T.; Takimura, T.; Sekine, R.; Kelly, V.P.; Kamata, K.; Sakamoto, K.; Nishimura, S.; Yokoyama, S.
Structural snapshots of the KMSKS loop rearrangement for amino acid activation by bacterial tyrosyl-tRNA synthetase
J. Mol. Biol.
346
105-117
2005
Escherichia coli (P0AGJ9), Escherichia coli
Iraha, F.; Oki, K.; Kobayashi, T.; Ohno, S.; Yokogawa, T.; Nishikawa, K.; Yokoyama, S.; Sakamoto, K.
Functional replacement of the endogenous tyrosyl-tRNA synthetase-tRNATyr pair by the archaeal tyrosine pair in Escherichia coli for genetic code expansion
Nucleic Acids Res.
38
3682-3691
2010
Saccharomyces cerevisiae, Methanocaldococcus jannaschii, Escherichia coli (P0AGJ9), Escherichia coli
Venkat, S.; Gregory, C.; Gan, Q.; Fan, C.
Biochemical characterization of the lysine acetylation of tyrosyl-tRNA synthetase in Escherichia coli
ChemBioChem
18
1928-1934
2017
Escherichia coli (P0AGJ9), Escherichia coli
Skupinska, M.; Stepniak, P.; Letowska, I.; Rychlewski, L.; Barciszewska, M.; Barciszewski, J.; Giel-Pietraszuk, M.
Natural compounds as inhibitors of tyrosyl-tRNA synthetase
Microb. Drug Resist.
23
308-320
2017
Staphylococcus aureus, Escherichia coli (P0AGJ9), Escherichia coli, Pseudomonas aeruginosa (Q9HWP3), Pseudomonas aeruginosa, Pseudomonas aeruginosa ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1 (Q9HWP3)
EXTERNAL LINKS (specific for EC-Number 6.1.1.1)
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
