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ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
primary binding pocket structure, overview
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
ATP + L-glutamate + tRNAGln
AMP + diphosphate + glutamyl-tRNAGln
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
ATP + L-glutamine + tRNAGln in unfractionated tRNA
?
-
-
-
-
?
additional information
?
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
wild-type tRNA, and var-AGGUtRNA, mechanism of the difference in the binding affinity of endogenous tRNAGln to the enzyme caused by noninterface nucleotides in variable loop, overview
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
primary binding pocket structure, overview
-
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + glutamyl-tRNAGln
-
-
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + glutamyl-tRNAGln
activity also with Gln-RS, EC 6.1.1.18, mutant C229R
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
r
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
-
-
-
r
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
specific tRNA-dependent amino acid recognition involves Asp66, Tyr211, and Phe233, which interact with A76 of tRNAGln and glutamine
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
two-step reaction: 1. recognition of appropriate amino acid by the enzyme and formation of an enzyme-bound mixed anhydride, the aminoacyl-AMP, under release of diphosphate, 2. transfer of the activated amino acid to the CCA end of the cognate tRNA to form aminoacyl-tRNA and AMP, both steps are tRNA-dependent
-
r
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
long-range intramolecular signaling in a tRNA synthetase complex
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
the enzyme is electrostatically optimized for binding of its cognate substrates
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
a two-step reaction
-
-
r
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
a two-step reaction, with a distinct role in induced-fit for Glu73
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
-
analysis of domain functions in enzyme-substrate interactions, overview
-
-
?
additional information
?
-
ternary complexed GlnRS bound to tRNAGln and the Gln-AMP analogue is catalytically active and has undergone the first step of the aminoacylation reaction
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-
?
additional information
?
-
-
ternary complexed GlnRS bound to tRNAGln and the Gln-AMP analogue is catalytically active and has undergone the first step of the aminoacylation reaction
-
-
?
additional information
?
-
-
wild-type GlnRS catalyzes Glu-tRNAGln synthesis 1000000fold less efficiently than the cognate reaction
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-
?
additional information
?
-
wild-type GlnRS catalyzes Glu-tRNAGln synthesis 1000000fold less efficiently than the cognate reaction
-
-
?
additional information
?
-
GlnRS has adetectable tRNA-acylation activity for its D-amino acid substrate
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-
?
additional information
?
-
-
GlnRS has adetectable tRNA-acylation activity for its D-amino acid substrate
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-
?
additional information
?
-
-
several noncognate tRNAs stimulate ATP-diphosphate exchange
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-
?
additional information
?
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-
lack of tRNA-independent diphosphate exchange
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-
?
additional information
?
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-
lack of tRNA-independent diphosphate exchange
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-
?
additional information
?
-
-
dimethyl sulfoxide stimulates the charging of several noncognate tRNA's with glutamine
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-
?
additional information
?
-
-
tRNA binding triggers aminoacyl-adenylate formation and diphosphate exchange
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-
?
additional information
?
-
-
importance of the acceptor binding domain for accurate recognition of tRNA
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-
?
additional information
?
-
-
conformational changes are induced by tRNAGln binding not by binding of tRNAGlu
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?
additional information
?
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-
phylogenetic analysis
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?
additional information
?
-
-
structure function analysis, overview
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-
?
additional information
?
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-
eukaryotic GlnRS evolves from GluRS by gene duplication and horizontally transfers to bacteria
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-
?
additional information
?
-
-
GlnRS forms a 1:1 molar complex with tRNAGln
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-
?
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0.26
L-glutamine
pH 7.5, 22Ā°C, wild-type enzyme
240
L-glutamate
mutant C229R GlnRS, with tRNAGln
0.0001
tRNAGln in unfractionated tRNA
-
-
-
additional information
additional information
-
0.04
ATP
-
wild-type enzyme, strain UT172
0.11
ATP
-
wild-type enzyme, pH 7.2, 37Ā°C
0.13
ATP
-
mutant D66E, pH 7.2, 37Ā°C
0.25
ATP
-
mutant Y211F/F233Y, pH 7.2, 37Ā°C
0.32
ATP
-
mutant Y211L, pH 7.2, 37Ā°C
0.42
ATP
-
mutant F233Y, pH 7.2, 37Ā°C
0.54
ATP
-
mutant F233D, pH 7.2, 37Ā°C
0.59
ATP
-
mutant D66F, pH 7.2, 37Ā°C
0.735
ATP
-
mutant Y211S, pH 7.2, 37Ā°C
0.74
ATP
-
mutant Y211F, pH 7.2, 37Ā°C
0.75
ATP
-
mutant F233L, pH 7.2, 37Ā°C
660
ATP
-
reaction with tRNA(2'H)Gln
0.11
Gln
-
Gln
0.19
Gln
-
wild-type enzyme, strain UT172
0.26
Gln
-
reaction with wild-type tRNAGln
1.43
Gln
-
reaction with mutant tRNAGln G36U
17.8
Gln
-
reaction with mutant tRNAGln U35A
0.05
L-glutamine
-
mutant F90L, pH 7.2, 37Ā°C
0.06
L-glutamine
-
truncated mutant, pH 7.2, 37Ā°C
0.07
L-glutamine
-
mutant Y240E, pH 7.2, 37Ā°C
0.114
L-glutamine
-
wild-type enzyme, pH 7.2, 37Ā°C
0.118
L-glutamine
-
mutant Y211L, pH 7.2, 37Ā°C
0.12
L-glutamine
-
mutant Y240G, pH 7.2, 37Ā°C
0.19
L-glutamine
-
wild-type enzyme, pH 7.2, 37Ā°C
0.21
L-glutamine
mutant C229R GlnRS, with tRNAGln
0.53
L-glutamine
-
mutant F233Y, pH 7.2, 37Ā°C
0.58
L-glutamine
-
mutant Y211F/F233Y, pH 7.2, 37Ā°C
0.76
L-glutamine
-
mutant D66F, pH 7.2, 37Ā°C
2.02
L-glutamine
-
mutant D66E, pH 7.2, 37Ā°C
2.21
L-glutamine
-
mutant F233L, pH 7.2, 37Ā°C
6.05
L-glutamine
-
mutant Y211S, pH 7.2, 37Ā°C
7.05
L-glutamine
-
mutant Y211F, pH 7.2, 37Ā°C
7.76
L-glutamine
-
mutant F233D, pH 7.2, 37Ā°C
10.4
L-glutamine
-
reaction with tRNA(2'H)Gln
22.3
L-glutamine
-
pH 7.2, 37Ā°C, recombinant mutant E73Q
34.9
L-glutamine
-
pH 7.2, 37Ā°C, recombinant mutant E34Q
45
L-glutamine
-
pH 7.2, 37Ā°C, recombinant mutant E34D
46.3
L-glutamine
-
pH 7.2, 37Ā°C, recombinant mutant E34A
0.000019
tRNAGln
-
wild-type enzyme, strain UT172
0.31
tRNAGln
-
wild-type tRNAGln
additional information
additional information
kinetics, tRNA substrate binding: calculation of the enthalpic and entropic contributions to the binding free energy with the molecular mechanics-Poisson-Boltzmann/surface area method, the entropic difference plays an important role in the difference in binding free energies, overview
-
additional information
additional information
-
no KM value for L-glutamate with the wild-type enzyme due to no saturation, kinetics of wild-type and mutant enzymes, overview. Kinetics of extended-loop GlnRS mutants, overview
-
additional information
additional information
no KM value for L-glutamate with the wild-type enzyme due to no saturation, kinetics of wild-type and mutant enzymes, overview. Kinetics of extended-loop GlnRS mutants, overview
-
additional information
additional information
-
Km value of mutant enzymes
-
additional information
additional information
-
single turnover kinetics, and steady-state kinetics of recombinant mutant enzymes, overview
-
additional information
additional information
-
kinetics of the mutant enzyme compared to wild-type GlnRS, EC 6.1.1.18, overview
-
additional information
additional information
kinetics of the mutant enzyme compared to wild-type GlnRS, EC 6.1.1.18, overview
-
additional information
additional information
-
pre-steady-state kinetics, negative allosteric coupling, overview
-
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0.046
L-glutamate
pH 7.5, 22Ā°C, wild-type enzyme
0.00041
L-glutamate
mutant C229R GlnRS, with tRNAGln
additional information
ATP
2.94
ATP
-
mutant F233Y, pH 7.2, 37Ā°C
2.94
ATP
-
mutant Y211F, pH 7.2, 37Ā°C
2.94
L-glutamine
-
mutant F233L, pH 7.2, 37Ā°C
3 - 6
L-glutamine
-
mutant F233Y, pH 7.2, 37Ā°C
3 - 6
L-glutamine
-
wild-type enzyme, pH 7.2, 37Ā°C
3.2
L-glutamine
pH 7.5, 22Ā°C, wild-type enzyme
6.08
L-glutamine
-
mutant Y211F, pH 7.2, 37Ā°C
0.016
ATP
-
mutant D66F, pH 7.2, 37Ā°C
0.042
ATP
-
mutant F233D, pH 7.2, 37Ā°C
0.14
ATP
-
mutant Y211S, pH 7.2, 37Ā°C
0.32
ATP
-
mutant Y211L, pH 7.2, 37Ā°C
0.46
ATP
-
mutant Y211F/F233Y, pH 7.2, 37Ā°C
1.51
ATP
-
mutant Y211F, pH 7.2, 37Ā°C
2.47
ATP
-
mutant F233Y, pH 7.2, 37Ā°C
2.75
ATP
-
mutant F233L, pH 7.2, 37Ā°C
2.8
ATP
-
wild-type enzyme, pH 7.2, 37Ā°C
6.27
ATP
-
mutant D66E, pH 7.2, 37Ā°C
0.0025
L-glutamine
mutant C229R GlnRS, with tRNAGln
0.004
L-glutamine
-
pH 7.2, 37Ā°C, recombinant mutant E73Q
0.014
L-glutamine
-
mutant D66F, pH 7.2, 37Ā°C
0.034
L-glutamine
-
pH 7.2, 37Ā°C, recombinant mutant E34D
0.04
L-glutamine
-
reaction with tRNA(2'H)Gln
0.05
L-glutamine
-
mutant F233D, pH 7.2, 37Ā°C
0.065
L-glutamine
-
pH 7.2, 37Ā°C, recombinant mutant E34Q
0.082
L-glutamine
-
mutant Y211S, pH 7.2, 37Ā°C
0.14
L-glutamine
-
pH 7.2, 37Ā°C, recombinant mutant E34A
0.4
L-glutamine
-
mutant Y211L, pH 7.2, 37Ā°C
0.55
L-glutamine
-
mutant Y211F/F233Y, pH 7.2, 37Ā°C
0.7
L-glutamine
-
mutant D66E, pH 7.2, 37Ā°C
1.4
L-glutamine
-
mutant Y240E, pH 7.2, 37Ā°C
1.48
L-glutamine
-
mutant Y211F, pH 7.2, 37Ā°C
1.62
L-glutamine
-
mutant F233L, pH 7.2, 37Ā°C
2
L-glutamine
-
mutant F90L, pH 7.2, 37Ā°C
2.62
L-glutamine
-
wild-type enzyme, pH 7.2, 37Ā°C
3.02
L-glutamine
-
mutant F233Y, pH 7.2, 37Ā°C
3.4
L-glutamine
-
mutant Y240G, pH 7.2, 37Ā°C
4.7
L-glutamine
-
wild-type enzyme and truncated mutant, pH 7.2, 37Ā°C
additional information
ATP
-
mutant D66E, pH 7.2, 37Ā°C
additional information
additional information
-
-
-
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C229R
site-directed mutagenesis, transplanting the conserved arginine residue from glutamyl-tRNA synthetase, EC 6.1.1.17, to glutaminyltRNA synthetase improves the KM of GlnRS for noncognate glutamate
C229R/Q255I
site-directed mutagenesis, comparison of mutant activity with glutamate and glutamine to charge tRNAGln to the wild-type activity, the mutant shows no activity with L-Gln, but weakly with L-Glu
C229R/Q255I/S227A/F233Y
site-directed mutagenesis, comparison of mutant activity with glutamate and glutamine to charge tRNAGln to the wild-type activity, the mutant shows no activity with L-Gln, but activity with L-Glu
D486R/L488Q
the double mutant causes a relaxed tRNA anticodon specificity
D81Q
site-diretced mutagenesis, the mutant has and increased, inverted stereospecificity. D81Q is predicted to lead to a rotated ligand backbone and an increased, not a decreased L-Tyr preference
E222K
site-directed mutagenesis, mutational structure-function study, the residue is part of the invariant Hub, the mutation leads to mischarging and affected cognate tRNAGln recognition
F90L
site-directed mutagenesis, mutational structure-function study, the residue is part of the connection in the active site network, the mutant shows increased Glu recognition in vitro and in vivo
Q255I
site-directed mutagenesis, mutational structure-function study, the residue is part of the invariant Hub, the mutation leads to reduced specificity for cognate Gln recognition and increased Glu recognition
R260Q
site-diretced mutagenesis, mutating Arg260 to the homologous but neutral Gln does not reduce the L-GlnAMP preference, instead, the mutation produces a change in the DELTADELTAG value that is much smaller than the wild-type free energy component
R30A
site-directed mutagenesis, comparison of mutant activity with glutamate and glutamine to charge tRNAGln to the wild-type activity, the mutant shows no activity with L-Glu
R30K
site-directed mutagenesis, comparison of mutant activity with glutamate and glutamine to charge tRNAGln to the wild-type activity, the mutant shows weak activity with L-Glu
R341A
site-directed mutagenesis, mutational structure-function study, the residue is part of the Hub common to all liganded complex, the mutation affects anticodon recognition
Y211H
site-directed mutagenesis, mutational structure-function study, the residue is part of the connection in the quaternary cognate-complex, the mutants shows slow solvation dynamics in the active site
Y240E/G
site-directed mutagenesis, mutational structure-function study, the residue is part of the Hub common to ligand-free and quaternary cognate-complex, the mutant shows increased Glu recognition in vitro and in vivo
A29X
-
site-directed mutagenesis
cGluGlnRS
-
a chimeric protein, consisting of the catalytic domain of GluRS and the anticodon-binding domain of GlnRS, is constructed
D235A
-
saturation mutagenesis, only little complementation of glnS-deficient strain
D66E
-
saturation mutagenesis, 18fold increased Km for glutamine, decreased turnover
D66F
-
saturation mutagenesis, highly increased Km for glutamine, 1200fold decrease in activity
D66G
-
saturation mutagenesis, only little complementation of glnS-deficient strain
D66H
-
saturation mutagenesis, only little complementation of glnS-deficient strain
D66R
-
saturation mutagenesis, only little complementation of glnS-deficient strain
E323A
-
site-directed mutagenesis, the mutation produces small but consistent 2 to 3fold improvements in glutamine-binding affinity compared to the wild-type enzyme
E34A
-
site-directed mutagenesis, the mutant shows highly increased Km and reduced kcat and activity compared to the wild-type enzyme
E34D
-
site-directed mutagenesis, the mutant shows highly increased Km and reduced kcat and activity compared to the wild-type enzyme
E34Q
-
site-directed mutagenesis, the mutant shows highly increased Km and reduced kcat and activity compared to the wild-type enzyme
E73A
-
site-directed mutagenesis, the mutant shows highly increased Km and reduced kcat and activity compared to the wild-type enzyme
E73Q
-
site-directed mutagenesis, the mutant shows highly increased Km and reduced kcat and activity compared to the wild-type enzyme, product release remains the rate-limiting step in E73Q
F233D
-
saturation mutagenesis, highly increased Km for glutamine, 3700fold decrease in activity
F233L
-
saturation mutagenesis, 19fold increased Km for glutamine, decreased turnover
F233Y
-
saturation mutagenesis, increased Km for glutamine, increased turnover
F90L
-
site-directed mutagenesis, active site mutant, 5fold improved glutamic acid recognition in vitro, in vivo the mutant shows a 40% reduced growth rate, partial complementation of an enzyme-deficient strain
K194A
-
site-directed mutagenesis, the mutation perturbs the dissociation constant in ATP binding
K401A
-
site-directed mutagenesis, the mutant shows reduced kcat compared to the wild-type enzyme
L136A
-
site-directed mutagenesis, the mutation perturbs the dissociation constant in ATP binding
N320A
-
site-directed mutagenesis, the mutation produces small but consistent 2 to 3fold improvements in glutamine-binding affinity compared to the wild-type enzyme
N336A
-
site-directed mutagenesis, the mutation removes contact with the ribose at U38, but does not significantly influence glutamine affinity
N370A
-
site-directed mutagenesis, the mutation removes contact with the base of U38, but does not significantly influence glutamine affinity
Q318A
-
site-directed mutagenesis, the mutation produces small but consistent 2 to 3fold improvements in glutamine-binding affinity compared to the wild-type enzyme
Q517A
-
site-directed mutagenesis, the mutant shows reduced kcat compared to the wild-type enzyme
R410A
-
site-directed mutagenesis, the mutation removes contact with the base of C34, but does not significantly influence glutamine affinity
R520A
-
site-directed mutagenesis, the mutant shows reduced kcat compared to the wild-type enzyme
R545A
-
site-directed mutagenesis, the mutant shows reduced kcat compared to the wild-type enzyme
T316A
-
site-directed mutagenesis, the mutation produces small but consistent 2 to 3fold improvements in glutamine-binding affinity compared to the wild-type enzyme
T547A
-
site-directed mutagenesis, the mutant shows reduced kcat compared to the wild-type enzyme
Y211F
-
saturation mutagenesis, 60fold increased Km for glutamine, decreased turnover
Y211F/F233Y
-
saturation mutagenesis, increased Km for glutamine, about 6fold decreased activity
Y211G
-
saturation mutagenesis, only little complementation of glnS-deficient strain
Y211L
-
saturation mutagenesis, unaffected Km for glutamine, decreased turnover
Y211S
-
saturation mutagenesis, 1700fold decrease in activity
Y240E
-
site-directed mutagenesis, active site mutant, 5fold improved glutamic acid recognition in vitro, partial complementation of an enzyme-deficient strain
Y240G
-
site-directed mutagenesis, active site mutant, 3fold improved glutamic acid recognition in vitro, partial complementation of an enzyme-deficient strain
R341A
-
site-directed mutagenesis, the mutation deletes a hydrogen bond made with the O4 moiety of the U35 base
R341A
-
site-directed mutagenesis, the mutation removes contact with the base of U35, but does not significantly influence glutamine affinity
additional information
-
cumulative replacement of other primary binding site residues than Cys229 in GlnRS, with those of GluRS, only slightly improves the ability of the GlnRS active site to accommodate glutamate. Introduction of 22 amino acid replacements and one deletion, including substitution of the entire primary binding site and two surface loops adjacent to the region disrupted in C229R, improves the capacity of Escherichia coli GlnRS to synthesize misacylated Glu-tRNAGln by 16000fold. This hybrid enzyme recapitulates the function of misacylating GluRS enzymes found in organisms that synthesize Gln-tRNAGln by an alternative pathway, overview
additional information
cumulative replacement of other primary binding site residues than Cys229 in GlnRS, with those of GluRS, only slightly improves the ability of the GlnRS active site to accommodate glutamate. Introduction of 22 amino acid replacements and one deletion, including substitution of the entire primary binding site and two surface loops adjacent to the region disrupted in C229R, improves the capacity of Escherichia coli GlnRS to synthesize misacylated Glu-tRNAGln by 16000fold. This hybrid enzyme recapitulates the function of misacylating GluRS enzymes found in organisms that synthesize Gln-tRNAGln by an alternative pathway, overview
additional information
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deletion mutants with C-terminal truncations and N-terminal truncations. A C-terminal deletion mutant exhibits sharp reduction in the specificity constant. Reduced stability of some of these mutants
additional information
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strain HAPPY101 allows plasmid-mediated expression of detrimental GlnRS mutants, which cannot complement the chromosomal glnS deletion in Escherichia coli strain X3R2
additional information
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3 misacylating mutant enzymes with reduced ability to discriminate between cognate and noncognate base pairs at position 3-70
additional information
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temperature-sensitive mutant enzyme, no change in affinity for glutamine
additional information
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construction of a truncated enzyme form, partial complementation of an enzyme-deficient strain, reduced growth rate in vivo
additional information
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construction of a chimeric glutamyl:glutaminyl-tRNA synthetase, cGluGlnRS, consisting of the catalytic domain of the GluRS and the anti-codon binding domain of the GlnRS. The chimeric mutant shows detectable glutamylation activity with Escherichia coli tRNAGlu and is capable of complementing a ts-GluRS strain at non-permissive temperatures. The GlnRS anticodon-binding domain in cGluGlnRS enhances kcat for glutamylation, interaction analysis, overview
additional information
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the engineered mutant hybrid C229R Gln-RS, EC 6.1.1.18, shows activity with L-glutamine or L-glutamate and tRNAGln like the nondiscriminating enzyme, EC 6.1.1.24. Introduction of 22 amino acid replacements and one deletion, including substitution of the entire primary binding site and two surface loops adjacent to the region disrupted in the mutant C229R, improves the capacity of the mutant enzyme to synthesize misacylated Glu-tRNAGln by 16000fold, overview
additional information
the engineered mutant hybrid C229R Gln-RS, EC 6.1.1.18, shows activity with L-glutamine or L-glutamate and tRNAGln like the nondiscriminating enzyme, EC 6.1.1.24. Introduction of 22 amino acid replacements and one deletion, including substitution of the entire primary binding site and two surface loops adjacent to the region disrupted in the mutant C229R, improves the capacity of the mutant enzyme to synthesize misacylated Glu-tRNAGln by 16000fold, overview
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