Information on EC 6.1.1.18 - Glutamine-tRNA ligase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
6.1.1.18
-
RECOMMENDED NAME
GeneOntology No.
Glutamine-tRNA ligase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ATP + L-glutamine + tRNAGln = AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln = AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
active site residues are Tyr240 and Phe90
-
ATP + L-glutamine + tRNAGln = AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
binding of ATP and of tRNAGln induces conformational changes that change the interaction of the enzyme with the cognate tRNA, crucial for substrate recognition and selectivity
-
ATP + L-glutamine + tRNAGln = AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
aminoacylation reaction mechanism, substrate binding sites, Tyr211 and a water molecule are responsible for recognition of both hydrogen atoms of the nitrogen of glutamine sidechain, which is esstial for substrate recognition
-
ATP + L-glutamine + tRNAGln = AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
structurefunction relationship, overview
-, P56926
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Aminoacylation
-
-
-
-
Aminoacylation
-
-
Aminoacylation
-
-
esterification
-
-
-
-
esterification
-
-
esterification
-
-
PATHWAY
KEGG Link
MetaCyc Link
Aminoacyl-tRNA biosynthesis
-
Metabolic pathways
-
tRNA charging
-
SYSTEMATIC NAME
IUBMB Comments
L-Glutamine:tRNAGln ligase (AMP-forming)
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
GlnRS
-
-
-
-
Glutamine translase
-
-
-
-
Glutamine--tRNA ligase
-
-
-
-
Glutamine-tRNA synthetase
-
-
-
-
glutaminyl tRNA synthetase
-
-
Glutaminyl-transfer ribonucleate synthetase
-
-
-
-
Glutaminyl-transfer RNA synthetase
-
-
-
-
Glutaminyl-tRNA synthetase
-
-
-
-
Glutaminyl-tRNA synthetase
-
-
Glutaminyl-tRNA synthetase
P00962
-
Glutaminyl-tRNA synthetase
-
-
glutaminyltRNA synthetase
P00962
-
Synthetase, glutaminyl-transfer ribonucleate
-
-
-
-
Vegetative specific protein H4
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9075-59-6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
gene glnS
SwissProt
Manually annotated by BRENDA team
K12 strain carrying the glnS structural gene on plasmid pBR322
-
-
Manually annotated by BRENDA team
misacylating mutants
-
-
Manually annotated by BRENDA team
purified recombinant wild-type and mutant enzymes
-
-
Manually annotated by BRENDA team
strain HAPPY101, plasmid-mediated expression of detrimental GlnRS mutants, which cannot complement the chromosomal glnS deletion in Escherichia coli strain X3R2
-
-
Manually annotated by BRENDA team
strains BT3213 and KL2576
-
-
Manually annotated by BRENDA team
strains UT172 and X3R2, and deletion mutants with C-terminal truncations and N-terminal truncations
-
-
Manually annotated by BRENDA team
temperature-resistant strain KL301 and temperature-sensitive mutant
-
-
Manually annotated by BRENDA team
enzyme forms a macromolecular protein complex
-
-
Manually annotated by BRENDA team
purified recombinant His-tagged enzyme expressed in Escherichia coli
-
-
Manually annotated by BRENDA team
strain PAO1
-
-
Manually annotated by BRENDA team
full-length and truncated enzyme form lacking the NH2-terminal domain
-
-
Manually annotated by BRENDA team
wild-type and mutant enzyme
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
negative allosteric coupling plays a key role in enforcing the selective RNA-amino acid pairing at the heart of the genetic code
evolution
-
the architecture of the GlnRS RNP has differentiated over evolutionary time to maintain glutamine-binding affinity at a weak level, and provides strong evidence for long-distance communication
additional information
-
generation of a comprehensive mapping of intramolecular communication in the glutaminyl-tRNA synthetase:tRNAGln complex, interaction analysis, detailed overview. Distinct coupling amplitudes for glutamine binding and aminoacyl-tRNA formation on the enzyme, respectively, implying the existence of multiple signaling pathways. Signaling from binding of the tRNA inner elbow, overview. Seven protein contacts with the distal tRNA vertical arm each weaken glutamine binding affinity across distances up to 40 A
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
P00962
primary binding pocket structure, overview
-
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + glutamyl-tRNAGln
show the reaction diagram
P00962
-, activity also with Gln-RS, EC 6.1.1.18, mutant C229R
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
r
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
r
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-, P56926
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
-
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
P00962
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
P00962
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
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
show the reaction diagram
-
tRNA substrate from bovine liver
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
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
show the reaction diagram
-
long-range intramolecular signaling in a tRNA synthetase complex
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
the enzyme is electrostatically optimized for binding of its cognate substrates
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
a two-step reaction
-
-
r
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
a two-step reaction, with a distinct role in induced-fit for Glu73
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
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
show the reaction diagram
-
analysis of domain functions in enzyme-substrate interactions, overview
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
P00962
primary binding pocket structure, overview
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
-
-
-
?
ATP + L-glutamine + tRNAGln(CUG)
AMP + diphosphate + L-glutaminyl-tRNAGln(CUG)
show the reaction diagram
-, P56926
-
-
-
?
ATP + L-glutamine + tRNAGln(UUG)
AMP + diphosphate + L-glutaminyl-tRNAGln(UUG)
show the reaction diagram
-, P56926
-
-
-
?
additional information
?
-
-
glutamine-dependent ATP-diphosphate exchange
-
-
-
additional information
?
-
-
several noncognate tRNAs stimulate ATP-diphosphate exchange
-
-
-
additional information
?
-
-
lack of tRNA-independent diphosphate exchange
-
-
-
additional information
?
-
-
dimethyl sulfoxide stimulates the charging of several noncognate tRNA's with glutamine
-
-
-
additional information
?
-
-
tRNA binding triggers aminoacyl-adenylate formation and diphosphate exchange
-
-
-
additional information
?
-
-
importance of the acceptor binding domain for accurate recognition of tRNA
-
-
-
additional information
?
-
-
conformational changes are induced by tRNAGln binding not by binding of tRNAGlu
-
?
additional information
?
-
-
phylogenetic analysis
-
?
additional information
?
-
-
the enzyme interacts with the apoptosis signal-regulating kinase ASK1, which involves the active sites of the enzymes and inhibits AKS1, the association is mediated and enhanced by glutamine, it is inhibited by Fas ligation, the enzyme inhibits ASK1-induced apoptosis
-
?
additional information
?
-
-
structure function analysis, 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
-
-
-
additional information
?
-
-, P56926
the Deinococcus radiodurans GlnRS is a structural hybrid between conventional GlnRS and AdT, EC 6.3.5.6, structurefunction relationship, the Yqey domain is involved in tRNAGln recognition and plays the role of an affinity enhancer of GlnRS for tRNAGln acting only in cis, overview
-
-
-
additional information
?
-
-
eukaryotic GlnRS evolves from GluRS by gene duplication and horizontally transfers to bacteria
-
-
-
additional information
?
-
P00962
wild-type GlnRS catalyzes Glu-tRNAGln synthesis 1000000fold less efficiently than the cognate reaction
-
-
-
additional information
?
-
-
GlnRS forms a 1:1 molar complex with tRNAGln
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + L-glutamate + tRNAGln
AMP + diphosphate + glutamyl-tRNAGln
show the reaction diagram
P00962
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
r
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
r
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-, P56926
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
P00962
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
P00962
-
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutaminyl-tRNAGln
show the reaction diagram
-
long-range intramolecular signaling in a tRNA synthetase complex
-
-
?
ATP + L-glutamine + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
phylogenetic analysis
-
?
additional information
?
-
-
the enzyme interacts with the apoptosis signal-regulating kinase ASK1, which involves the active sites of the enzymes and inhibits AKS1, the association is mediated and enhanced by glutamine, it is inhibited by Fas ligation, the enzyme inhibits ASK1-induced apoptosis
-
?
additional information
?
-
-
eukaryotic GlnRS evolves from GluRS by gene duplication and horizontally transfers to bacteria
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ATP
-
binding of ATP induces conformational changes that change the interaction of the enzyme with the cognate tRNA
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Co2+
-
can partially replace Mg2+ in activation of ATP-diphosphate exchange
Mg2+
-
Km: 8.9 mM; optimal concentration: 25 mM; required
Mg2+
-
optimal MgCl2/ATP ratio is 2-5 at 2 mM ATP in aminoacylation; required
Mg2+
P00962
;
Mg2+
-
required
Mn2+
-
can replace Mg2+ in activation with 42% efficiency
Mn2+
-
can replace Mg2+ in activation of aminoacylation
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
4-Methyleneglutamine
-
-
5'-O-[N-(L-glutaminyl)sulfamoyl] adenosine
-
; i.e. QSI, competitive to glutamine
5'-O-[N-(L-glutaminyl)sulfamoyl] adenosine
-
i.e. QSI, glutaminyl-adenylate analogue, competitive to glutamine
5,5'-dithiobis(2-nitrobenzoate)
-
-
Br-
-
0.1 M, 50% inhibition
Cl-
-
0.2 M, 50% inhibition
Glutamic acid 4-hydroxamate
-
-
glutaminol adenylate
-
; competitive to glutamine
glutaminol adenylate methyl phosphate ester
-
; competitive to glutamine
glutaminyl-beta-ketophosphonate-adenosine
-
i.e. Gln-KPA, selective, competitive inhibition of GlnRS, contrast, Gln-KPA inhibits GlnRS by binding competitively but weakly at two distinct sites on the enzyme, the glutamine and the AMP modules of Gln-KPA, connected by the beta-ketophosphonate linker, cannot bind GlnRS simultaneously, and that one Gln-KPA molecule binds the AMP-binding site of GlnRS through its AMP module, whereas another Gln-KPA molecule binds the glutamine-binding site through its glutamine module, mechanism, overview
-
glutamyl-beta-ketophosphonate-adenosine
-
i.e. Glu-KPA, competitive inhibition, non-cognate, binds weakly at one site on the monomeric enzyme
-
I-
-
0.06 M, 50% inhibition
L-Glutamic acid
-
competitive inhibition of the wild-type and mutant enzymes
p-hydroxymercuribenzoate
-
-
S-Carbamoylcysteine
-
-
S-Carbamoylserine
-
-
tRNA
-
above 0.6 mg/ml
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
putrescine
-
can partially replace Mg2+ in activation, with 32% efficiency
-
spermidine
-
can partially replace Mg2+ in activation, with 12% efficiency
spermine
-
can partially replace Mg2+ in activation, with 35% efficiency
spermine
-
significant stimulation of ATP-diphosphate exchange and aminoacylation in presence of limited MgCl2 concentrations, cannot totally replace Mg2+
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.038
-
ATP
-
full-length enzyme
0.04
-
ATP
-
wild-type enzyme, strain UT172
0.11
-
ATP
-
wild-type enzyme, pH 7.2, 37C
0.111
-
ATP
-
-
0.13
-
ATP
-
mutant D66E, pH 7.2, 37C
0.21
-
ATP
-
-
0.25
-
ATP
-
mutant Y211F/F233Y, pH 7.2, 37C
0.26
-
ATP
-
truncated enzyme
0.32
-
ATP
-
mutant Y211L, pH 7.2, 37C
0.42
-
ATP
-
mutant F233Y, pH 7.2, 37C
0.54
-
ATP
-
mutant F233D, pH 7.2, 37C
0.59
-
ATP
-
mutant D66F, pH 7.2, 37C
0.735
-
ATP
-
mutant Y211S, pH 7.2, 37C
0.74
-
ATP
-
mutant Y211F, pH 7.2, 37C
0.75
-
ATP
-
mutant F233L, pH 7.2, 37C
660
-
ATP
-
reaction with tRNA(2'H)Gln
0.028
-
Gln
-
full-length enzyme
0.11
-
Gln
-
truncated enzyme
0.11
-
Gln
-
Gln
0.15
-
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
240
-
L-glutamate
P00962
mutant C229R GlnRS, with tRNAGln
0.05
-
L-glutamine
-
mutant F90L, pH 7.2, 37C
0.06
-
L-glutamine
-
truncated mutant, pH 7.2, 37C
0.07
-
L-glutamine
-
mutant Y240E, pH 7.2, 37C
0.114
-
L-glutamine
-
wild-type enzyme, pH 7.2, 37C
0.118
-
L-glutamine
-
mutant Y211L, pH 7.2, 37C
0.12
-
L-glutamine
-
mutant Y240G, pH 7.2, 37C
0.19
-
L-glutamine
-
wild-type enzyme, pH 7.2, 37C
0.21
-
L-glutamine
P00962
mutant C229R GlnRS, with tRNAGln
0.26
-
L-glutamine
P00962
pH 7.5, 22C, wild-type enzyme
0.53
-
L-glutamine
-
mutant F233Y, pH 7.2, 37C
0.58
-
L-glutamine
-
mutant Y211F/F233Y, pH 7.2, 37C
0.76
-
L-glutamine
-
mutant D66F, pH 7.2, 37C
2.02
-
L-glutamine
-
mutant D66E, pH 7.2, 37C
2.21
-
L-glutamine
-
mutant F233L, pH 7.2, 37C
6.05
-
L-glutamine
-
mutant Y211S, pH 7.2, 37C
7.05
-
L-glutamine
-
mutant Y211F, pH 7.2, 37C
7.76
-
L-glutamine
-
mutant F233D, pH 7.2, 37C
10.4
-
L-glutamine
-
reaction with tRNA(2'H)Gln
22.3
-
L-glutamine
-
pH 7.2, 37C, recombinant mutant E73Q
34.9
-
L-glutamine
-
pH 7.2, 37C, recombinant mutant E34Q
45
-
L-glutamine
-
pH 7.2, 37C, recombinant mutant E34D
0.000019
-
tRNAGln
-
wild-type enzyme, strain UT172
0.0002
-
tRNAGln
-
-
0.0017
-
tRNAGln
-
full-length enzyme and truncated enzyme
0.31
-
tRNAGln
-
wild-type tRNAGln
0.0001
-
tRNAGln in unfractionated tRNA
-
-
-
46.3
-
L-glutamine
-
pH 7.2, 37C, recombinant mutant E34A
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, 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
-, P56926
effect of the isolated Yqey domain on the kinetic properties of GlnRS, overview
-
additional information
-
additional information
P00962
kinetics of the mutant enzyme compared to wild-type GlnRS, EC 6.1.1.18, overview; 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
-
pre-steady-state kinetics, negative allosteric coupling, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.016
-
ATP
-
mutant D66F, pH 7.2, 37C
0.042
-
ATP
-
mutant F233D, pH 7.2, 37C
0.14
-
ATP
-
mutant Y211S, pH 7.2, 37C
0.32
-
ATP
-
mutant Y211L, pH 7.2, 37C
0.46
-
ATP
-
mutant Y211F/F233Y, pH 7.2, 37C
1.51
-
ATP
-
mutant Y211F, pH 7.2, 37C
2.47
-
ATP
-
mutant F233Y, pH 7.2, 37C
2.75
-
ATP
-
mutant F233L, pH 7.2, 37C
2.8
-
ATP
-
wild-type enzyme, pH 7.2, 37C
2.94
-
ATP
-
mutant F233Y, pH 7.2, 37C; mutant Y211F, pH 7.2, 37C
6.27
-
ATP
-
mutant D66E, pH 7.2, 37C
0.00041
-
L-glutamate
P00962
mutant C229R GlnRS, with tRNAGln
0.046
-
L-glutamate
P00962
pH 7.5, 22C, wild-type enzyme
0.0025
-
L-glutamine
P00962
mutant C229R GlnRS, with tRNAGln
0.004
-
L-glutamine
-
pH 7.2, 37C, recombinant mutant E73Q
0.014
-
L-glutamine
-
mutant D66F, pH 7.2, 37C
0.034
-
L-glutamine
-
pH 7.2, 37C, recombinant mutant E34D
0.04
-
L-glutamine
-
reaction with tRNA(2'H)Gln
0.05
-
L-glutamine
-
mutant F233D, pH 7.2, 37C
0.065
-
L-glutamine
-
pH 7.2, 37C, recombinant mutant E34Q
0.082
-
L-glutamine
-
mutant Y211S, pH 7.2, 37C
0.14
-
L-glutamine
-
pH 7.2, 37C, recombinant mutant E34A
0.4
-
L-glutamine
-
mutant Y211L, pH 7.2, 37C
0.55
-
L-glutamine
-
mutant Y211F/F233Y, pH 7.2, 37C
0.7
-
L-glutamine
-
mutant D66E, pH 7.2, 37C
1.4
-
L-glutamine
-
mutant Y240E, pH 7.2, 37C
1.48
-
L-glutamine
-
mutant Y211F, pH 7.2, 37C
1.62
-
L-glutamine
-
mutant F233L, pH 7.2, 37C
2
-
L-glutamine
-
mutant F90L, pH 7.2, 37C
2.62
-
L-glutamine
-
wild-type enzyme, pH 7.2, 37C
2.94
-
L-glutamine
-
mutant F233L, pH 7.2, 37C
3
6
L-glutamine
-
mutant F233Y, pH 7.2, 37C; wild-type enzyme, pH 7.2, 37C
3.02
-
L-glutamine
-
mutant F233Y, pH 7.2, 37C
3.2
-
L-glutamine
P00962
pH 7.5, 22C, wild-type enzyme
3.4
-
L-glutamine
-
mutant Y240G, pH 7.2, 37C
4.7
-
L-glutamine
-
wild-type enzyme and truncated mutant, pH 7.2, 37C
3.3
-
tRNAGln
-
-
additional information
-
ATP
-
mutant D66E, pH 7.2, 37C
6.08
-
L-glutamine
-
mutant Y211F, pH 7.2, 37C
additional information
-
additional information
-
-
-
additional information
-
additional information
-
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0013
-
5'-O-[N-(L-glutaminyl)sulfamoyl] adenosine
-
versus glutamine
0.0013
-
5'-O-[N-(L-glutaminyl)sulfamoyl] adenosine
-
pH 7.0, 37C
0.00028
-
glutaminol adenylate
-
versus glutamine
0.01
-
glutaminol adenylate methyl phosphate ester
-
versus glutamine
0.65
-
glutaminyl-beta-ketophosphonate-adenosine
-
pH 7.2, 37C, versus L-glutamine
-
2.8
-
glutamyl-beta-ketophosphonate-adenosine
-
pH 7.2, 37C, versus L-glutamine
-
17
-
L-Glutamic acid
-
truncated mutant, pH 7.2, 37C
21
-
L-Glutamic acid
-
mutant Y240E, pH 7.2, 37C
29
-
L-Glutamic acid
-
mutant F90L, pH 7.2, 37C
47
-
L-Glutamic acid
-
mutant Y240G, pH 7.2, 37C
96
-
L-Glutamic acid
-
wild-type enzyme, pH 7.2, 37C
additional information
-
additional information
-
inhibition kinetics
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.1074
-
-
-
1.28
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
activities of recombinant mutant enzymes
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.2
-
-
ATP-diphosphate exchange
7
-
-
assay at
7.2
-
-, P56926
assay at
7.2
-
-
assay at
7.5
-
-
assay at
7.5
-
P00962
assay at; assay at
8.5
-
-
aminoacylation
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
22
-
P00962
assay at room temperature; assay at room temperature
25
-
-
assay at
37
-
-, P56926
assay at
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
localization of cytoplasmic GlnRS in mitochondrial Gln-tRNA synthesis
Manually annotated by BRENDA team
-
localization of cytoplasmic GlnRS in mitochondrial Gln-tRNA synthesis. Saccharomyces cerevisiae imports the cytosolic pathway for Gln-tRNA synthesis into the mitochondrion
Manually annotated by BRENDA team
-
enzyme DNA sequence contains a lysine-rich nuclear targeting sequence motif
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / LMG 4051 / NBRC 15346 / NCIMB 9279 / R1 / VKM B-1422)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
65400
71200
-
sucrose density gradient sedimentation
68500
-
-
electrophoresis of the native enzyme in polyacrylamide gels of various concentrations
138000
-
-
gel filtration
additional information
-
-
structural similarities in glutaminyl-tRNA synthetase and methionyl-tRNA synthetase outside the respective dinucleotide-fold domains
additional information
-
-
primary structure
additional information
-
-
primary structure
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
monomer
-
1 * 91000, SDS-PAGE
monomer
-
1 * 69000, SDS-PAGE
monomer
-
1 * 64500, SDS-PAGE
monomer
-
1 * 64200, recombinant His6-tagged GlnRS, SDS-PAGE
monomer
-
1 * 64000
additional information
-
enzyme is part of a high molecular mass aminoacyl-tRNA synthetase complex, which has a coherent structure, that can be visualized by electron microscopy
additional information
-
targeting into the multienzyme complex is mediated by the C-terminal catalytic domain
additional information
-
structure function analysis, overview
additional information
-
molecular dynamics simulations on wild-type tRNA, var-AGGUtRNA, and tRNA-GlnRS complexes, overview
additional information
-, P56926
the enzyme possesses a C-terminal extension of 215 residues appending the anticodon-binding domain, the Yqey domain, which constitutes a paralog of the Saccharomyces cerevisiae Yqey protein, structure-function relationship, the Yqey domain is involved in tRNAGln recognition and plays the role of an affinity enhancer of GlnRS for tRNAGln acting only in cis, overview
additional information
-
GlnRS structure networks, detection method development for accounting side chain interactions, yet providing a global view of the ligand-induced conformational changes, and understand allosteric changes mediated by the binding of ligands, usage of crystal structures: PDB IDs 1nyl, 1qtq, 1o0b, and 1o0c, overview
additional information
-
determination of glutamyl/glutaminyl-tRNA synthetase domain sequences in the proteosome of the organism, classification of proteins into homologous groups by determination and validation of Markov clusters of homologous subsequences, MACHOS, for structure-function analysis, method evaluation, overview
additional information
-
long-range signal propagation from the tRNA anticodon is dynamically driven, whereas shorter pathways are mediated by induced-fit rearrangements, structure modelling, overview
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
purified recombinant full-length GlnRS grown in microbatch in the presence of PEG 3350, X-ray diffraction structure determination and analysis at 2.3 A resolution, molecular replacement
-, P56926
vapour-diffusion method. Orthorombic crystals are obtained that belong to space group P2(1)2(1)2(1) and diffract to 2.3 A resolution
-
2.5 A resolution; structure of the enzyme with its cognate glutaminyl-tRNA and ATP; the entire anticodon loop provides essential sites for glutaminyl tRNA synthetase discrimination among tRNA molecules
-
2.8 A resolution; structure of the enzyme with its cognate glutaminyl-tRNA and ATP
-
analysis of the crystal structure of GlnRS-tRNAGln complex bound to the glutaminyl adenylate analogue 5'-O-[N-(L-Gln)sulfamoyl] adenosine
-
cocrystallization of the purified enzyme with tRNAGln and inhibitor QSI, X-ray diffraction structure determination at 2.4 A resolution and analysis
-
crystal structure of three misacylating mutants of Escherichia coli glutaminyl-tRNA synthetase complexed with tRNAGln and ATP
-
crystals of the GlnRS-tRNA(2'H)Gln complex bound to the ATP analog AMPCPP and glutamine are grown by microseeding with crystals of the GlnRS-tRNAGln-ATP ternary complex. Crystals grew in 1-2 weeks by vapor diffusion over a reservoir containing 2 M ammonium sulfate, 10 mM Pipes, pH 7.5, 10 mM MgCl2 and 2 mM DTT
-
detailed structural comparison between Met-tRNA synthetase and Gln-tRNA synthetase
-
GlnRS-tRNAGln complex, 6.6 mg/ml protein in 10 mM PIPES, pH 7.5, 10 mM MgCl2, and 1.8-5.4 mM tRNA. The tRNA/analog solution is then mixed with equal volumes of a 6.3 mg/ml solution of GlnRS, containing 5mM PIPES, pH 7.0, and 5 mM 2-mercaptoethanol, X-ray diffraction structure determination and analysis at 2.6 A resolution; purified recombinant GlnRS C229R-tRNAGln complex, a protein solution containing 6.3mg/ml GlnRS prepared in 5 mM PIPES, pH 7.0, 5 mM 2-mercaptoethanol, is mixed with the tRNAGln solution, X-ray diffraction structure determination and analysis at 2.6 A resolution
P00962
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
40
-
-
strain KL301 enzyme is stable, temperature-sensitive mutant enzyme loses about 70% of its activity
additional information
-
-
several noncognate tRNAs protect the enzyme against thermal inactivation
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, 20 mM potassium phosphate, pH 7.5, 50 mM KCl, 1 mM DTT, 50% glycerol, stable for 6 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant His-tagged chimeric mutant enzyme from strain BL21(DE3) or the temperature sensitive strain JP1449(DE3) by nickel affinity chromatography
-
recombinant His-tagged GlnRS mutants from strain BL21-DE3 by nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21
P00962
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression of full length and C-terminal truncated GlnRS, lacking the the Yqey domain, and of the isolated Yqey protein, in Escherichia coli strain ER2566
-, P56926
overexpression in Escherichia coli
-
expression of His-tagged GlnRS mutants in strain BL21-DE3
-
expression of the His-tagged chimeric mutant enzyme in Escherichia coli strain BL21(DE3) or the temperature sensitive strain JP1449(DE3)
-
gene glnS, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21
P00962
the vector pET28a is used, Escherichia coli JP1449DE3 cells are used
-
overexpression of various Myc-tagged enzyme mutants in human embryonic kidney 293 cells
-
DNA sequence determination and analysis, enzyme contains a N-terminal lysine-rich sequence KPKKKKEK, that may function as a nuclear targetng signal as well as in regulation of gene expression, since the 8 amino acid peptide derived from the motif interacts with DNA and changes the DNA molecule form from B to Z form
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
A29X
-
site-directed mutagenesis
C229R
P00962
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
P00962
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
P00962
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
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
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
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
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
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
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
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
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
R30A
P00962
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
P00962
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
R341A
-
site-directed mutagenesis, the mutation deletes a hydrogen bond made with the O4 moiety of the U35 base; site-directed mutagenesis, the mutation removes contact with the base of U35, but does not significantly influence glutamine affinity
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
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
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
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
L136A
-
site-directed mutagenesis, the mutation perturbs the dissociation constant in ATP binding
additional information
-
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
-
strain HAPPY101 allows plasmid-mediated expression of detrimental GlnRS mutants, which cannot complement the chromosomal glnS deletion in Escherichia coli strain X3R2
additional information
-
3 misacylating mutant enzymes with reduced ability to discriminate between cognate and noncognate base pairs at position 3-70
additional information
-
temperature-sensitive mutant enzyme, no change in affinity for glutamine
additional information
-
construction of a truncated enzyme form, partial complementation of an enzyme-deficient strain, reduced growth rate in vivo
additional information
-
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
P00962
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; 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
Y240G
-
site-directed mutagenesis, active site mutant, 3fold improved glutamic acid recognition in vitro, partial complementation of an enzyme-deficient strain
additional information
-
a deletion mutant comprising only the C-terminal catalytic domain is targeted into the multienzyme complex, while a deletion mutant comprising only the N-terminal domain is not
additional information
-
truncated enzyme form lacking the NH2-terminal domain, with modest increase in Km value for glutamine and ATP and no difference in kcat for aminoacylation or Km for tRNA
additional information
-
mutant enzyme with increased Km for glutamine
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
urea induces equilibrium denaturation of glutaminyl-tRNA synthetase, existence of a stable intermediate state at around 2 M urea, existence of an induced molten globule state in a large multidomain protein which is separated from the native and the denatured protein by high activation energy barriers
-