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Information on EC 6.1.1.3 - threonine-tRNA ligase and Organism(s) Escherichia coli and UniProt Accession P0A8M3

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The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
Synonyms
threonyl-trna synthetase, thrrs, thr-trna synthetase, mitochondrial threonyl-trna synthetase, threonyl trna synthetase, threonine-trna ligase, threonyl-transfer ribonucleic acid synthetase, bathrrs, apthrrs-1, apthrrs-2, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Synthetase, threonyl-transfer ribonucleate
-
Threonine translase
-
Threonine--tRNA ligase
-
Threonine-transfer ribonucleate synthetase
-
threonyl tRNA synthetase
-
Threonyl-ribonucleic synthetase
-
Threonyl-transfer ribonucleate synthetase
-
Threonyl-transfer ribonucleic acid synthetase
-
Threonyl-transfer RNA synthetase
-
Synthetase, threonyl-transfer ribonucleate
Threonine translase
Threonine--tRNA ligase
Threonine-transfer ribonucleate synthetase
Threonyl-ribonucleic synthetase
Threonyl-transfer ribonucleate synthetase
Threonyl-transfer ribonucleic acid synthetase
Threonyl-transfer RNA synthetase
Threonyl-tRNA synthetase
ThrRS
additional information
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
ATP + L-threonine + tRNAThr = AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
esterification
Aminoacylation
esterification
Aminoacylation
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
L-threonine:tRNAThr ligase (AMP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY hide
9023-46-5
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + hydroxynorvaline + tRNAThr
AMP + diphosphate + hydroxynorvalyl-tRNAThr
show the reaction diagram
10-70% less active than with L-threonine
-
?
ATP + L-serine + tRNAThr
AMP + diphosphate + L-seryl-tRNAThr
show the reaction diagram
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
ATP + 3-hydroxynorvaline + tRNAThr
AMP + diphosphate + 3-hydroxynorvalyl-tRNAThr
show the reaction diagram
-
the specificity constant kcat/KM for beta-hydroxynorvaline is only 20-30fold less than that of cognate threonine, amino acid activation is the potential rate-limiting step of b3-hydroxynorvaline aminoacylation
-
-
?
ATP + L-serine + tRNASer
AMP + diphosphate + L-seryl-tRNASer
show the reaction diagram
-
-
-
-
?
ATP + L-serine + tRNAThr
AMP + diphosphate + L-seryl-tRNAThr
show the reaction diagram
-
very low activity with the wild-type enzyme
-
?
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
ATP + L-threonine + tRNAThr
AMP + diphosphate + L-threonyl-tRNAThr
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
cofactor binding structure, modeling
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ni2+
the enzyme can bind one Ni2+ per subunit, binding of nickel inhibits the oxidation of enzyme residue Cys182 by H2O2 or air
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
borrelidin
H2O2
oxidation of ThrRS by H2O2 causes editing defects and Ser misincorporation at Thr codons due to oxidation of Cys182, zinc or nickel ions inhibit C182 oxidation by hydrogen peroxide. Reducing the oxidized ThrRS with DTT or sodium arsenite (NaAsO2) recovers the editing activity, cysteine residue C182 is reversibly oxidized
operator mRNA domain 2
-
-
Zn2+
inhibits the editing reaction
(2S,3R)-2,3-diamino-N-(((E)-3-(6-aminopyrimidin-4-yl)-styryl)sulfonyl)butanamide
-
-
(2S,3R)-2-amino-3-hydroxy-N-((3-(1-oxoisoindolin-5-yl)-phenyl)sulfonyl)butanamide
-
-
(2S,3R)-2-amino-3-hydroxy-N-((3-(3-methyl-1H-indazol-5-yl)phenyl)sulfonyl)butanamide
-
-
(2S,3R)-2-amino-3-hydroxy-N-((4-phenoxyphenyl)sulfonyl)-butanamide
-
-
(2S,3R)-2-amino-3-hydroxy-N-methyl-N-((3-(1-oxoisoindolin-5-yl)phenyl)sulfonyl)butanamide
-
-
(2S,3R)-2-amino-N'-(3-(4-amino-2-chloroquinazolin-7-yl)-phenyl)-3-hydroxybutanehydrazide
-
-
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxy-4-methylpentanamide
-
-
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxybutanamide
-
-
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxypentanamide
-
-
(2S,3R)-2-amino-N-((3-(1-amino-3-chloroisoquinolin-6-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
-
-
(2S,3R)-2-amino-N-((3-(1-aminoisoquinolin-6-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
-
-
(2S,3R)-2-amino-N-((3-(2,4-diaminoquinazolin-7-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
-
-
(2S,3R)-2-amino-N-((3-(3-chloro-1H-indazol-5-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
-
-
(2S,3R)-2-amino-N-((3-(4-amino-2-chloroquinazolin-7-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
-
-
(2S,3R)-2-amino-N-((3-(4-amino-2-methylquinazolin-7-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
-
-
(2S,3R)-2-amino-N-((3-(4-aminoquinazolin-7-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
-
-
(2S,3R)-2-amino-N-((7-(6-aminopyrimidin-4-yl)naphthalen-2-yl)sulfonyl)-3-hydroxybutanamide
-
-
(2S,3R)-2-amino-N-(3-(4-amino-2-chloroquinazolin-7-yl)-benzyl)-3-hydroxybutanamide
-
-
(2S,3R)-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)sulfonyl)-2,3-dihydroxybutanamide
-
-
(2S,3R)-N-((7-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-2-amino-3-hydroxybutanamide
-
-
(2S,3R)-N-((7-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)naphthalen-2-yl)sulfonyl)-2-amino-3-hydroxybutanamide
-
-
5'-O-[N-(threonyl)-sulfamoyl] adenosine
-
-
borrelidin
-
-
hydrogen peroxide
-
oxidizes cysteine182 residue critical for editing, which leads to Ser-tRNAThr formation and protein mistranslation that impaired growth of Escherichia coli. Presence of major heat shock proteases is required to allow cell growth in medium containing serine and hydrogen peroxide, which suggests that the mistranslated proteins are misfolded
tert-butyl((2S,3R)-1-(3-(1H-indazol-5-yl)-benzenesulfonamido)-3-(tert-butoxy)-1-oxobutan-2-yl)-carbamate
-
-
threonyl-AMP
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
3-hydroxynorvaline enhances the ATPase function of the synthetic site, at a rate not increased by nonaminoacylatable tRNA
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.95 - 7
hydroxynorvaline
81.5 - 142
L-serine
0.101 - 0.18
L-threonine
0.00003 - 0.00043
tRNAThr
0.267 - 0.387
ATP
0.00003 - 0.897
L-threonine
0.00005 - 0.0015
tRNAThr
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
21 - 22
hydroxynorvaline
26 - 30
L-serine
2 - 37
L-threonine
42 - 90
ATP
0.64 - 90
L-threonine
0.05 - 0.7
tRNAThr
additional information
additional information
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
38.3
tRNAThr
-
pH 7.2, 37°C, ThrRS, aminoacylation with Thr
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0000034 - 0.006
borrelidin
0.05
(2S,3R)-2,3-diamino-N-(((E)-3-(6-aminopyrimidin-4-yl)-styryl)sulfonyl)butanamide
-
Ki above 0.05 mM with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0000077
(2S,3R)-2-amino-3-hydroxy-N-((3-(1-oxoisoindolin-5-yl)-phenyl)sulfonyl)butanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.000127
(2S,3R)-2-amino-3-hydroxy-N-((3-(3-methyl-1H-indazol-5-yl)phenyl)sulfonyl)butanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0365
(2S,3R)-2-amino-3-hydroxy-N-((4-phenoxyphenyl)sulfonyl)-butanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.05
(2S,3R)-2-amino-3-hydroxy-N-methyl-N-((3-(1-oxoisoindolin-5-yl)phenyl)sulfonyl)butanamide
-
Ki above 0.05 mM, with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.000136
(2S,3R)-2-amino-N'-(3-(4-amino-2-chloroquinazolin-7-yl)-phenyl)-3-hydroxybutanehydrazide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0000324
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxy-4-methylpentanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0000027
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxybutanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0000043
(2S,3R)-2-amino-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)-sulfonyl)-3-hydroxypentanamide
-
Ki above 0.05 mM, with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.00002
(2S,3R)-2-amino-N-((3-(1-amino-3-chloroisoquinolin-6-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.00009
(2S,3R)-2-amino-N-((3-(1-aminoisoquinolin-6-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0000107
(2S,3R)-2-amino-N-((3-(2,4-diaminoquinazolin-7-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.000052
(2S,3R)-2-amino-N-((3-(3-chloro-1H-indazol-5-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0000008 - 0.05
(2S,3R)-2-amino-N-((3-(4-amino-2-chloroquinazolin-7-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
0.0000172
(2S,3R)-2-amino-N-((3-(4-amino-2-methylquinazolin-7-yl)-phenyl)sulfonyl)-3-hydroxybutanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0000029
(2S,3R)-2-amino-N-((3-(4-aminoquinazolin-7-yl)phenyl)-sulfonyl)-3-hydroxybutanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0000027
(2S,3R)-2-amino-N-((7-(6-aminopyrimidin-4-yl)naphthalen-2-yl)sulfonyl)-3-hydroxybutanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.000329
(2S,3R)-2-amino-N-(3-(4-amino-2-chloroquinazolin-7-yl)-benzyl)-3-hydroxybutanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.05
(2S,3R)-N-(((E)-3-(6-aminopyrimidin-4-yl)styryl)sulfonyl)-2,3-dihydroxybutanamide
-
Ki above 0.05 mM, with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0000095
(2S,3R)-N-((7-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3,4-dihydroisoquinolin-2(1H)-yl)sulfonyl)-2-amino-3-hydroxybutanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0000024
(2S,3R)-N-((7-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)naphthalen-2-yl)sulfonyl)-2-amino-3-hydroxybutanamide
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.0000131
5'-O-[N-(threonyl)-sulfamoyl] adenosine
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
0.000182
tert-butyl((2S,3R)-1-(3-(1H-indazol-5-yl)-benzenesulfonamido)-3-(tert-butoxy)-1-oxobutan-2-yl)-carbamate
-
with L-serine as substrate, in 60 mM Tris, pH 7.6, 10 mM MgCl2, 20 mM KCl, temperature not specified in the publication
additional information
additional information
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.7
aminoacylation assay at
7.2
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
threonyl-tRNA synthetase (ThrRS) misactivates serine and utilizes an editing site cysteine (C182 in Escherichia coli) to hydrolyze Ser-tRNAThr. Hydrogen peroxide oxidizes C182, leading to SertRNAThr production and mistranslation of threonine codons as serine. C182 is oxidized to sulfenic acid by air, hydrogen peroxide, and hypochlorite. Air oxidation increases the Ser-tRNAThr level in the presence of elongation factor Tu. C182 forms a putative metal binding site with three conserved histidine residues (H73, H77, and H186). H73 and H186, but not H77, are critical for activating C182 for oxidation. Zinc or nickel ions inhibit C182 oxidation by hydrogen peroxide. Bacteria may use ThrRS editing to sense the oxidant levels in the environment. C182 oxidation modeling, overview. C182 is directly activated by H73 and H186 rather than by a metal ion. Chronic oxidative stress leads to ThrRS mistranslation in vivo
physiological function
malfunction
-
reactive oxygen species cause editing defect and misacylation by WT ThrRS. H2O2-induced Ser-tRNAThr formation causes protein mistranslation
physiological function
-
amino acid discrimination does not occur at the aminoacyl transfer step. pre-Transfer hydrolysis contributes to proofreading only when the rate of transfer is slowed significantly. Thus, the relative contributions of pre- and posttransfer editing in ThrRS are subject to modulation by the rate of aminoacyl transfer
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
150000
164000
-
PAGE at different gel concentrations
76000
-
2 * 76000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
dimer
additional information
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure, PDB code 1EVL, at 1.55 A resolution used for reaction mechanism analysis and molecular modeling
enzyme core DELTAN complexed with the essential operator domain mRNA, precipitation with 1.9 M ammonum sulfate in sodium cacodylate, pH 6.6, 4°C from solution containing 0.088 mM enzyme mutant DELTAN, 0.2 mM operator mRNA domain 2, 10 mM MgCl2, 10 mM substrate analogue 5'-O-(N-(L-threonyl)-sulfamoyl)adenosine, crystals are soaked in 25% v/v glycerol, X-ray diffraction structure determination at 3.6 A resolution and structure analysis
purified recombinant N-terminal part of the enzyme, i.e. N1 and N2 domains comprising residues 1-65 and 66-225 in one fragment, X-ray diffraction structure determination and analysis at 1.5 A resolution, structure modeling
X-ray structure determination at 2.9 A resolution, structure analysis of the wild-type enzyme and truncated mutant lamdaN in complex with L-threonine and L-serine
enzyme in complex with tRNAThr, hanging drop method, pH 6.5, solution containing ammonium acetate, ATP, and PEG 4000 as precipitant, X-ray diffraction structure determination at 2.9 A resolution and analysis
-
hanging drop vapor diffusion method, using 12% (w/v) PEG 4000, 21% (v/v) 2-methyl-2,4-pentanediol, and 0.1 M sodium citrate, pH 5.9
-
lamdaN-threonine-tRNA ligase complexed with Ser-AMS, X-ray diffraction structure determination at 1.65 A resolution and analysis
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C182A
site-directed mutagenesis, the mutation leads to loss of editing activity and to Ser misacylation to tRNAThr. C182A and C182S mutations reduce the kcat value of editing over 500fold
C182S
site-directed mutagenesis, the mutation leads to loss of editing activity and to Ser misacylation to tRNAThr. C182A and C182S mutations reduce the kcat value of editing over 500fold
D435A
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
D46E
site-directed mutagenesis, the mutant has a modest reduction in its aminoacylation activity compared to wild-type, and its post-transfer editing activity is abolished
D46E/H186G
site-directed mutagenesis, the mutant EcThrRS strongly supports growth of a yeast thrS deletion strain (ScDELTAthrS)
D46E/Y173F
site-directed mutagenesis, the mutant EcThrRS supports growth of a yeast thrS deletion strain (ScDELTAthrS)
D46E/Y173H
site-directed mutagenesis, the mutant does not support growth of a yeast thrS deletion strain (ScDELTAthrS)
D46E/Y173K
site-directed mutagenesis, the mutant does not support growth of a yeast thrS deletion strain (ScDELTAthrS)
D46E/Y173R
site-directed mutagenesis, the mutant does not support growth of a yeast thrS deletion strain (ScDELTAthrS)
D46E/Y173S
site-directed mutagenesis, the mutant does not support growth of a yeast thrS deletion strain (ScDELTAthrS)
D46R
site-directed mutagenesis, the mutant has a modest reduction in its aminoacylation activity compared to wild-type, and its post-transfer editing activity is abolished
E458D
site-directed mutagenesis, the sensitivity of the mutant enzyme to borrelidin is reduced markedly compared to wild-type, mutant shows decreased apparent rate constants
G459D
site-directed mutagenesis, the sensitivity of the mutant enzyme to borrelidin is reduced markedly compared to wild-type, mutant shows decreased apparent rate constants
H186A
site-directed mutagenesis, the mutant does not show oxidation of Cys182 by H2O2 and only partially by NaOCl
H186G
site-directed mutagenesis, the mutant EcThrRS strongly supports growth of a yeast thrS deletion strain (ScDELTAthrS)
H309A
site-directed mutagenesis, mutant shows highly increased Ki for inhibitor borrelidin compared to the wild-type enzyme
H337A
site-directed mutagenesis, mutant shows increased Ki for inhibitor borrelidin compared to the wild-type enzyme
H73A
site-directed mutagenesis, the mutant does not show oxidation of Cys182 by H2O2 and NaOCl
H77A
site-directed mutagenesis, the mutant shows oxidation of Cys182 by H2O2 and NaOCl
K136A
site-directed mutagenesis, the mutant EcThrRS supports growth of a yeast thrS deletion strain (ScDELTAthrS)
K136E
site-directed mutagenesis, the mutant does not support growth of a yeast thrS deletion strain (ScDELTAthrS)
K136R
site-directed mutagenesis, the mutant EcThrRS supports growth of a yeast thrS deletion strain (ScDELTAthrS)
L489W
site-directed mutagenesis, mutant has a reduced space of the hydrophobic cluster near the active site resulting in a 1500fold increase in Ki for inhibitor borrelidin compared to the wild-type enzyme
P296A
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
P296S
site-directed mutagenesis, mutant shows slightly increased Ki for inhibitor borrelidin compared to the wild-type enzyme
P335A
site-directed mutagenesis, mutant shows increased Ki for inhibitor borrelidin compared to the wild-type enzyme
P424K
site-directed mutagenesis, the sensitivity of the mutant enzyme to borrelidin is reduced markedly compared to wild-type, the mutant shows decreased apparent rate constants
P464A
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
R282A
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
S429A
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
T307A
site-directed mutagenesis, mutant shows increased Ki for inhibitor borrelidin compared to the wild-type enzyme
Y173D
site-directed mutagenesis, the mutant has a modest reduction in its aminoacylation activity compared to wild-type, and its post-transfer editing activity is abolished
Y173R
site-directed mutagenesis, the mutant has a modest reduction in its aminoacylation activity compared to wild-type, and its post-transfer editing activity is abolished
Y313A
site-directed mutagenesis, mutant shows a similar Ki for inhibitor borrelidin compared to the wild-type enzyme
D180A
-
charging of tRNAThr with serine, mutant is no longer able to rapidly deacetylate Ser-tRNAThr
D549A
-
modified interation of anticodon loop/C-ter domain, activity similar to the wild-type
E258K
-
modified interation with the superrepressor, no activity
E259K
-
modified interation with the superrepressor, unaltered activity
E600A
-
modified interation of anticodon loop/C-ter domain, 710fold increased activity
H73A
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme, and a 2fold higher rate of ATP consumption relative to the rate of Ser-tRNAThr synthesis
H73A/H309A
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme, and a 2fold higher rate of ATP consumption relative to the rate of Ser-tRNAThr synthesis
H73A/H77A
-
charging of tRNAThr with serine, mutant is no longer able to deacetylate Ser-tRNAThr
K246A
-
modified interation of acceptor stem and catalytic domain, 2.9fold increased activity
K249A
-
modified interation of acceptor stem and catalytic domain, 3.5fold increased activity
K577A
-
modified interation of anticodon loop/C-ter domain, 118fold increased activity
N324A
-
modified interation of cross-subunit contacts, 3.5fold increased activity
N502A
-
modified interation of cross-subunit contacts, 2.1fold increased activity
N575A
-
modified interation of anticodon loop/C-ter domain, 9.4fold increased activity
R349A
-
modified interation of cross-subunit contacts, 42fold increased activity
R583H
-
modified interation of anticodon loop/C-ter domain, no activity
R609A
-
modified interation of anticodon loop/C-ter domain, 35fold activity
S347A
-
modified interation of cross-subunit contacts, similar to the wild-type
S367A
-
modified interation of acceptor stem and catalytic domain, 11fold increased activity
W434Y
-
reduced activity, Trp434 is involved in conformational changes during substrate binding
Y205F
-
modified interation of acceptor stem and N-terminal domain, 7.7fold increased activity
Y219F
-
modified interation of acceptor stem and N-terminal domain, similar to the wild-type
Y348F
-
modified interation of cross-subunit contacts, 6.5fold increased activity
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60
-
12 min, incubation without substrates, 50% inactivation
additional information
-
threonine does not influence the rate of heat inactivation. ATP and tRNA labilize
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, stable for more than 18 months
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-type and mutant enzyme from Escherichia coli mutant by nickel affinity chromatography
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant non-tagged N1 and N2 domains comprising residues 1-65 and 66-225 in one fragment by ion exchange chromatography, ammonium sulfate fractionation, hydrophobic interaction chromatography, and ultrafiltration
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3)
metal chelate affinity column chromatography
-
recombinant His-tagged enzyme from strain IBPC6881, to homogeneity
-
recombinant His-tagged ThrRS by nickel affinity chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression of His-tagged wild-type and mutant enzyme in an enzyme-deficient Escherichia coli mutant, complementation analysis
expression of non-tagged N1 and N2 domains comprising residues 1-65 and 66-225 in one fragment in strain BL21
gene thrS, sequence comparisons, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
gene thrS, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
expression of His-tagged ThrRS
-
overexpression in strain IBPC6881 as His-tagged enzyme
-
overexpression of wild-type and mutants in the null mutant strain
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Freist, W.; Gauss, D.H.
Threonyl-tRNA synthetase.
Biol. Chem. Hoppe-Seyler
376
213-224
1995
Aesculus hippocastanum, Geobacillus stearothermophilus, Bos taurus, Saccharomyces cerevisiae, Oryctolagus cuniculus, Escherichia coli, Thermus thermophilus, Homo sapiens, Mus musculus, Rattus norvegicus, Saccharomyces pastorianus
Manually annotated by BRENDA team
Zheltonosova, J.; Melnikova, E.; Garber, M.; Reinbolt, J.; Kern, D.; Ehresmann, C.; Ehresmann, B.
Threonyl-tRNA synthetase from Thermus thermophilus. Purification and some structural and kinetic properties
Biochimie
76
71-77
1994
Escherichia coli, Thermus thermophilus, Thermus thermophilus HB8 / ATCC 27634 / DSM 579
Manually annotated by BRENDA team
Paetz, W.; Nass, G.
Biochemical and immunological characterization of threonyl-tRNA synthetase of two borrelidin-resistant mutants of Escherichia coli K12
Eur. J. Biochem.
35
331-337
1973
Escherichia coli
Manually annotated by BRENDA team
Hennecke, H.; B`ck, A.; Thomale, J.; Nass, G.
Threonyl-transfer ribonucleic acid synthetase from Escherichia coli. Subunit structure and genetic analysis of the structural gene by means of a mutated enzyme and of a specialized transducing lambda bacteriophage
J. Bacteriol.
131
943-950
1977
Escherichia coli, Escherichia coli overproducing
Manually annotated by BRENDA team
Bovee, M.L.; Pierce, M.A.; Francklyn, C.S.
Induced fit and kinetic mechanism of adenylation catalyzed by Escherichia coli threonyl-tRNA synthetase
Biochemistry
42
15102-15113
2003
Escherichia coli
Manually annotated by BRENDA team
Dock-Bregeon, A.; Sankaranarayanan, R.; Romby, P.; Caillet, J.; Springer, M.; Rees, B.; Francklyn, C.S.; Ehresmann, C.; Moras, D.
Transfer RNA-mediated editing in threonyl-tRNA synthetase. The class II solution to the double discrimination problem
Cell
103
877-884
2000
Escherichia coli
Manually annotated by BRENDA team
Sankaranarayanan, R.; Dock-Bregeon, A.C.; Romby, P.; Caillet, J.; Springer, M.; Rees, B.; Ehresmann, C.; Ehresmann, B.; Moras, D.
The structure of threonyl-tRNA synthetase-tRNA(Thr) complex enlightens its repressor activity and reveals an essential zinc ion in the active site
Cell
97
371-381
1999
Escherichia coli
Manually annotated by BRENDA team
Caillet, J.; Nogueira, T.; Masquida, B.; Winter, F.; Graffe, M.; Dock-Bregeon, A.C.; Torres-Larios, A.; Sankaranarayanan, R.; Westhof, E.; Ehresmann, B.; Ehresmann, C.; Romby, P.; Springer, M.
The modular structure of Escherichia coli threonyl-tRNA synthetase as both an enzyme and a regulator of gene expression
Mol. Microbiol.
47
961-974
2003
Escherichia coli
Manually annotated by BRENDA team
Sankaranarayanan, R.; Dock-Bregeon, A.C.; Rees, B.; Bovee, M.; Caillet, J.; Romby, P.; Francklyn, C.S.; Moras, D.
Zinc ion mediated amino acid discrimination by threonyl-tRNA synthetase
Nat. Struct. Biol.
7
461-465
2000
Escherichia coli (P0A8M3)
Manually annotated by BRENDA team
Torres-Larios, A.; Dock-Bregeon, A.C.; Romby, P.; Rees, B.; Sankaranarayanan, R.; Caillet, J.; Springer, M.; Ehresmann, C.; Ehresmann, B.; Moras, D.
Structural basis of translational control by Escherichia coli threonyl tRNA synthetase
Nat. Struct. Biol.
9
343-347
2002
Escherichia coli (P0A8M3), Escherichia coli
Manually annotated by BRENDA team
Ruan, B.; Bovee, M.L.; Sacher, M.; Stathopoulos, C.; Poralla, K.; Francklyn, C.S.; Soll, D.
A unique hydrophobic cluster near the active site contributes to differences in borrelidin inhibition among threonyl-tRNA synthetases
J. Biol. Chem.
280
571-577
2005
Archaeoglobus fulgidus, Halobacterium sp., Helicobacter pylori, Methanocaldococcus jannaschii, Saccharolobus solfataricus, Escherichia coli (P0A8M3), Escherichia coli
Manually annotated by BRENDA team
Dock-Bregeon, A.C.; Rees, B.; Torres-Larios, A.; Bey, G.; Caillet, J.; Moras, D.
Achieving error-free translation; the mechanism of proofreading of threonyl-tRNA synthetase at atomic resolution
Mol. Cell
16
375-386
2004
Escherichia coli (P0A8M3)
Manually annotated by BRENDA team
Zurek, J.; Bowman, A.L.; Sokalski, W.A.; Mulholland, A.J.
MM and QM/MM modeling of threonyl-tRNA synthetase: model testing and simulations
Struct. Chem.
15
405-414
2004
Escherichia coli (P0A8M3)
-
Manually annotated by BRENDA team
Minajigi, A.; Deng, B.; Francklyn, C.S.
Fidelity escape by the unnatural amino acid beta-hydroxynorvaline: an efficient substrate for Escherichia coli threonyl-tRNA synthetase with toxic effects on growth
Biochemistry
50
1101-1109
2011
Escherichia coli
Manually annotated by BRENDA team
Minajigi, A.; Francklyn, C.S.
Aminoacyl transfer rate dictates choice of editing pathway in threonyl-tRNA synthetase
J. Biol. Chem.
285
23810-23817
2010
Escherichia coli
Manually annotated by BRENDA team
Ling, J.; Soell, D.
Severe oxidative stress induces protein mistranslation through impairment of an aminoacyl-tRNA synthetase editing site
Proc. Natl. Acad. Sci. USA
107
4028-4033
2010
Escherichia coli, Escherichia coli MG1655
Manually annotated by BRENDA team
Teng, M.; Hilgers, M.T.; Cunningham, M.L.; Borchardt, A.; Locke, J.B.; Abraham, S.; Haley, G.; Kwan, B.P.; Hall, C.; Hough, G.W.; Shaw, K.J.; Finn, J.
Identification of bacteria-selective threonyl-tRNA synthetase substrate inhibitors by structure-based design
J. Med. Chem.
56
1748-1760
2013
Burkholderia thailandensis, Escherichia coli, Homo sapiens (P26639), Yersinia pestis
Manually annotated by BRENDA team
Bushnell, E.A.; Huang, W.; Llano, J.; Gauld, J.W.
Molecular dynamics investigation into substrate binding and identity of the catalytic base in the mechanism of threonyl-tRNA synthetase
J. Phys. Chem. B
116
5205-5212
2012
Escherichia coli (P0A8M3)
Manually annotated by BRENDA team
Li, M.; Zhang, J.; Liu, C.; Fang, B.; Wang, X.; Xiang, W.
Identification of borrelidin binding site on threonyl-tRNA synthetase
Biochem. Biophys. Res. Commun.
451
485-490
2014
Escherichia coli (P0A8M3), Escherichia coli
Manually annotated by BRENDA team
Zhou, X.L.; Chen, Y.; Fang, Z.P.; Ruan, Z.R.; Wang, Y.; Liu, R.J.; Xue, M.Q.; Wang, E.D.
Translational quality control by bacterial threonyl-tRNA synthetases
J. Biol. Chem.
291
21208-21221
2016
Mycoplasma capricolum, Escherichia coli (P0A8M3), Escherichia coli, Mesomycoplasma mobile (Q6KH76), Mesomycoplasma mobile ATCC 43663 (Q6KH76)
Manually annotated by BRENDA team
Dutta, S.; Nandi, N.
Classical molecular dynamics simulation of seryl tRNA synthetase and threonyl tRNA synthetase bound with tRNA and aminoacyl adenylate
J. Biomol. Struct. Dyn.
2018
1-23
2018
Escherichia coli (P0A8M3)
Manually annotated by BRENDA team
Wu, J.; Fan, Y.; Ling, J.
Mechanism of oxidant-induced mistranslation by threonyl-tRNA synthetase
Nucleic Acids Res.
42
6523-6531
2014
Escherichia coli (P0A8M3)
Manually annotated by BRENDA team