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Information on EC 6.1.1.17 - glutamate-tRNA ligase and Organism(s) Escherichia coli and UniProt Accession P04805
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6.1.1.17 glutamate-tRNA ligaseSpecify your search results
Word Map
- 6.1.1.17
- synthetases
- aminoacyl-trna
- aminoacylation
- glnrs
- trnagln
- ampa
-
ionotropic
-
excitatory
-
anticodon
-
metabotropic
- kainate
- glu-trnagln
-
amidotransferase
- n-methyl-d-aspartate
-
misacylated
-
glutamatergic
-
mischarging
-
trna-dependent
-
glutaminylation
-
non-nmda
-
noncognate
-
nmdars
-
transamidation
- aspartyl-trna
- gatcab
-
aarss
- metrs
- trnatyr
-
isoacceptors
- lysyl-trna
-
non-discriminating
-
misaminoacylation
-
alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate
-
ampa-type
- glutamate-1-semialdehyde
- arginyl-trna
-
asparaginyl-trna
-
anticodon-binding
-
glur2/3
- methionyl-trna
-
trna-binding
- drug development
-
tryptophanyl-trna
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
glurs, glutaminyl-trna synthetase, glutamyl-trna synthetase, glurs2, trna modifying enzyme, glurs1, glutamyl trna synthetase, discriminating glurs, glursat, d-glurs, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
GluRS
Glutamate--tRNA ligase
-
-
-
-
Glutamate-tRNA synthetase
-
-
-
-
Glutamic acid translase
-
-
-
-
Glutamic acid tRNA ligase
-
-
-
-
Glutamyl tRNA synthetase
-
-
-
-
Glutamyl-transfer ribonucleate synthetase
-
-
-
-
Glutamyl-transfer ribonucleic acid synthetase
-
-
-
-
Glutamyl-transfer RNA synthetase
-
-
-
-
Glutamyl-tRNA synthetase
P85
-
-
-
-
GluRS
-
-
-
-
Glutamyl-tRNA synthetase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + L-glutamate + tRNAGlu = AMP + diphosphate + L-glutamyl-tRNAGlu
aminoacylation reaction via a two-step mechanism involving a very unstable aa-AMP intermediate
ATP + L-glutamate + tRNAGlu = AMP + diphosphate + L-glutamyl-tRNAGlu
concerted mechanism
-
ATP + L-glutamate + tRNAGlu = AMP + diphosphate + L-glutamyl-tRNAGlu
ATP and tRNAGlu bind randomly to the free enzyme, whereas glutamate binds only to the ternary enzyme-tRNAGlu-ATP complex. After interconversion of the quarternary enzyme-substrate complex the end-products dissociate in the order: diphosphate, AMP, Glu-tRNA
-
ATP + L-glutamate + tRNAGlu = AMP + diphosphate + L-glutamyl-tRNAGlu
the major recognition element of the tRNAGlu is U at position 34
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
esterification
Aminoacylation
esterification
-
-
-
-
Aminoacylation
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
KEGG
-
-, -
SYSTEMATIC NAME
IUBMB Comments
L-glutamate:tRNAGlu ligase (AMP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY
9068-76-2
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + L-glutamate + tRNAAsp
AMP + diphosphate + L-glutamyl-tRNAAsp
-
adB gene encodes a truncated GluRS that lacks the C-terminal third of the protein and, consequently the anticodon binding domain. The YadB protein transfers Glu onto tRNAAsp. Neither tRNAGlu nor tRNAGln are substrates
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
?
ATP + Glu + tRNAAsp
AMP + diphosphate + L-glutamyl-tRNAAsp
-
the enzyme glutamylates the queuosine residue, a modified nucleoside at the wooble position of the tRNAASp QUC anticodon. The enzyme is not able to glutamylate tRNAAsp isolated from an Escherichia coli tRNA-guanosine transglycosylase minus strain deprived of the capacity to exchange guanosine 34 with queuosine
-
-
?
ATP + Glu + tRNAAsp
AMP + diphosphate + L-glutamyl-tRNAAsp
-
tRNAGlu is not used as substrate
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
r
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
-
r
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
existence of an enzyme-AMP-Glu intermediate in the aminoacylation reaction
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
specific modification of the 5-(methylaminomethyl)-2-thiouridine group in the anticodon of E. coli tRNAGlu by cyanogen bromide results in a 5fold decrease of the maximal rate of Glu-tRNAGlu synthesis, but unaffected rate of tRNAGlu-promoted ATP-diphosphate exchange
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
the major recognition element of the tRNAGlu is U at position 34, activity with wild-type and mutant tRNAs, overview
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
a two-step reaction
-
-
r
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
ERS recognizes the 2-thionyl group of 2-thio-5-methylaminomethyluridine in the first or wobble anticodon position of tRNAGlu, specific, though tenuous interaction, recognition determinants and mechanism, overview
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
wild-type enzyme and chimeric mutant cGluGlnRS, overview
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
analysis of domain functions in enzyme-substrate interactions, overview
-
-
?
additional information
?
-
efficient glutamylation demands optimal binding of substrates (ATP, tRNAGlu and L-glutamic acid) by GluRS. Binding of tRNAGlu induces conformational changes in GluRS that stimulates the binding of L-glutamic acid leading to the productive binding of ATP. L-glutamic acid is first activated by GluRS in presence of ATP to form the adenylate complex. This is followed by the catalytic step where the acceptor stem of tRNAGlu is glutamylated
-
-
?
additional information
?
-
-
efficient glutamylation demands optimal binding of substrates (ATP, tRNAGlu and L-glutamic acid) by GluRS. Binding of tRNAGlu induces conformational changes in GluRS that stimulates the binding of L-glutamic acid leading to the productive binding of ATP. L-glutamic acid is first activated by GluRS in presence of ATP to form the adenylate complex. This is followed by the catalytic step where the acceptor stem of tRNAGlu is glutamylated
-
-
?
additional information
?
-
-
the identity of tRNAGlu is determined by the bases set U34,U35, C36, A37, G1.C72, U2.A71, U11.A24, U13.G22..A46, and DELTA47, if this set is transplanted to tRNAAsp in addition to C4.G69 and C12.G23..C9, the tRNAAsp is a substrate for the enzyme, while tRNAGlu modification at the bases of the determinant set results in reduced activity
-
?
additional information
?
-
-
the enzyme also catalyzes ATP-diphosphate exchange
-
-
?
additional information
?
-
-
thermal stability and structural analysis of tRNA substrates, overview
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
r
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
-
-
-
r
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
-
wild-type enzyme and chimeric mutant cGluGlnRS, overview
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Zn2+
dispensability of zinc and the putative zinc-binding domain in bacterial glutamyl-tRNA synthetase, overview. The association of enzyme GluRS with tRNAGlu but not with ATP is sensitive to perturbations of the zinc binding domain. Natively zinc-bound Ec-GluRS does not require zinc to be active
Mg2+
Mn2+
-
can partially replace Mg2+ in activation, maximal efficiency at 5 mM is 71% of Mg2+-activation
Zinc
Zinc
-
no direct interaction of the zinc atom with tRNA. It may stabilize a region of polypeptide chain which is involved in acceptor stem binding
Zinc
-
enzyme contains one zinc atom strongly bound, which is essential for its native conformation and its catalytic acitivity
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
5'-O-[N-(L-glutamyl)sulfamoyl]adenosine
i.e. glutamyl-sulfamoyl-adenosine or Glu-AMS, tRNAGlu increases the affinity of glutamyl-tRNA synthetase for its inhibitor glutamyl-sulfamoyl-adenosine, an analogue of the aminoacylation reaction intermediate glutamyl-AMP, thermodynamics of the enzyme-inhibitor interactions, overview. A significant entropic contribution for the interactions between Glu-AMS and GluRS in the absence of tRNA or in the presence of the cognate tRNAGlu or of the non-cognate tRNAPhe is indicated. The large negative enthalpy is the dominant contribution to DELTAGb in the absence of tRNA. The affinity of GluRS for Glu-AMS is not altered in the presence of the non-cognate tRNAPhe, but the dissociation constant Kd is decreased 50fold in the presence of tRNAGlu. Presence of an H-bond between Glu-AMS and the 3'-OH oxygen of the 3'-terminal ribose of tRNAGlu in the Glu-AMS/GluRS/tRNAGlu complex, molecular dynamics study
5'-O-(N-(L-glutamyl)-sulfamoyl)adenosine
-
potent competitive with respect to glutamic acid
glutaminyl-beta-ketophosphonate-adenosine
-
i.e. Gln-KPA, competitive inhibition, non-cognate, binds at one site on the monomeric enzyme
glutamyl-beta-ketophosphonate-adenosine
-
i.e. Glu-KPA, selective, competitive inhibition of GluRS, binds at one site on the monomeric enzyme
N6-Benzoyl-L-glutamyl AMP
-
specific for glutamyl-tRNA synthetase, does not inhibit glutaminyl-tRNA synthetase
additional information
-
ERS aminoacylation of tRNAGlu is inhibited by the tRNA fragments, RNA-protein interactions, ERS binding of minihelixGlu and fragments ASLGlu and ASLGlu-s2U34, overview
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
kinetics, diverse tRNAGlu mutants
-
additional information
additional information
-
Km for diverse modified forms of tRNAAsp and tRNAGlu
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
activity with diverse modified forms of tRNAAsp and tRNAGlu
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0000028
5'-O-[N-(L-glutamyl)sulfamoyl]adenosine
pH and temperature not specified in the publication
2.9
glutaminyl-beta-ketophosphonate-adenosine
-
pH 7.2, 37°C, versus L-glutamate
0.018
glutamyl-beta-ketophosphonate-adenosine
-
pH 7.2, 37°C, versus L-glutamate
additional information
additional information
-
kinetics of inhibitory tRNA fragments, overview
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
35 - 55
-
35°C: about 70% of maximal activity, 55°C: about 60% of maximal activity
additional information
additional information
microcalorimetry analysis at 20°C, 30°C and 37°C, thermodynamics in presence and absence of tRNAGlu and inhibitor Glu-AMS and non-cognate aa-AMP
additional information
-
microcalorimetry analysis at 20°C, 30°C and 37°C, thermodynamics in presence and absence of tRNAGlu and inhibitor Glu-AMS and non-cognate aa-AMP
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
the structures of the active sites of bacterial and mammalian GluRSs differ significantly
evolution
-
many bacterial GluRS are capable of recognizing two tRNA substrates: tRNAGlu and tRNAGln, e.g. GluRS from such as Bacillus subtilis, Thermosynechococcus elongatus, and Mycobacterium tuberculosis. In bacteria such as Escherichia coli and Thermus thermophilus that possess glutaminyl-tRNA synthetase (GlnRS), the cognate aminoacylating enzyme for tRNAGln, GluRS exclusively glutamylates tRNAGlu. tRNA-GluRS interaction in bacteria is also associated with phylum-specific idiosyncrasies, structure-function analysis, overview
additional information
additional information
homology structure modeling using structures of GluRSs identified from Burkholderia thailandensis and Thermosynechococcus elongatus, Uniprot IDs Q2SX36 and Q8DLI5, respectively, as search template, molecular docking
additional information
-
homology structure modeling using structures of GluRSs identified from Burkholderia thailandensis and Thermosynechococcus elongatus, Uniprot IDs Q2SX36 and Q8DLI5, respectively, as search template, molecular docking
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
46000
-
1 * 56000 (catalytic subunit) + 1 * 46000 (no detectable enzymatic activity, protective function), SDS-PAGE
56000
56000
-
1 * 56000 (catalytic subunit) + 1 * 46000 (no detectable enzymatic activity, protective function), SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
-
1 * 56000 (catalytic subunit) + 1 * 46000 (no detectable enzymatic activity, protective function), SDS-PAGE
monomer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified Gly-GluRS K236E/E328A, hanging drop vapour diffusion method, mixing of 0.001 ml of 20 mg/ml protein in 20 mM HEPES, pH 7.5, 50 mM NaCl, 10 mM 2-mercaptoethanol, and 50 mM ZnCl2, with 0.0012 ml of crystallization solution containing 0.1 M MOPS/HEPES-Na, pH 7.7, 0.02 M each of L-Glu.Na, DL-Ala, DL-Lys-HCl, Gly and DL-Ser, 14% w/v PEG 8000, 22% v/v ethylene glycol, 2 weeks, method optimization, X-ray diffraction structure determination and analysis at 3.5 A resolution, molecular replacement
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
H129Q
-
mutants encoding GluRS variants altered in the 98C-138C segment. Thermosensitive mutants H129Q, H131Q, H132Q and C138S. Mutants without glutamyl-tRNA synthetase activity: C100S, C125S. In the mutants C98S and H127Q the activity is 10fold lower than in cells overproducing the wild-type enzyme or the variants H129Q, H131Q, H132Q, and C138S
K236E/E328A
-
by mapping crystal contacts of the homologous GluRS from Bacillus thailandensis, PDB ID 4g6z, onto the Escherichia coli GluRS sequence, two surface residues are identified that might be hindering crystallization attempts. Accordingly, these two residues are mutated and crystallization of the double mutant is attempted
additional information
additional information
construction of constructed a chimaeric protein, cGluGlnRS, consisting of the catalytic domain, GluRS, and the anticodon binding domain of Escherichia coli GlnRS. cGluGlnRS shows detectable activity of glutamylation of Escherichia coli tRNAGlu and is capable of complementing an Escherichia coli temperature-sensitive GluRS strain at non-permissive temperatures. Both cGluGlnRS and N-terminal residues 1-314 of GluRS bind Escherichia coli tRNAglu with native GluRS-like affinity, suggesting that the anticodon-binding domain in cGluGlnRS enhances kcat for glutamylation. The kcat value of cGluGlnRS is approx. 500fold lower than that of GluRS, whereas the Km value is only moderately higher at the same solution conditions
additional information
-
construction of constructed a chimaeric protein, cGluGlnRS, consisting of the catalytic domain, GluRS, and the anticodon binding domain of Escherichia coli GlnRS. cGluGlnRS shows detectable activity of glutamylation of Escherichia coli tRNAGlu and is capable of complementing an Escherichia coli temperature-sensitive GluRS strain at non-permissive temperatures. Both cGluGlnRS and N-terminal residues 1-314 of GluRS bind Escherichia coli tRNAglu with native GluRS-like affinity, suggesting that the anticodon-binding domain in cGluGlnRS enhances kcat for glutamylation. The kcat value of cGluGlnRS is approx. 500fold lower than that of GluRS, whereas the Km value is only moderately higher at the same solution conditions
additional information
generation of pZBD-chimeras of Ec-GluRS, four chimeric versions with partly replaced zinc-binding motif, pZBD. In the first chimera [Ec(Te)-GluRS], the pZBD of Ec-GluRS is replaced by the corresponding pZBD from Thermosynechococcus elongatus GluRS (Te-GluRS) whose structure is devoid of a bound Zn2+ despite containing the modified ZB-motif CxCxnYx3H. In the second chimera [Ec(EQRS)-GluRS] the pZBD of Ec-GluRS is replaced by the corresponding pZBD from Escherichia coli Glu-QRS (Ec-EQRS) whose zinc-bound structure contains the ZB-motif CxCxnYx3C. In the third chimera [Ec(Bt)-GluRS], the pZBD of Ec-GluRS is replaced by a 19-residue stretch from the pZBD of Bt-GluRS which contains a disrupted zinc binding motif CxMx20Yx3W and whose structure is devoid of a bound Zn2+ ion. The 19-residue stretch starts from the residue preceding the fourth zinc-co-ordinating cysteine residue in Ec-GluRS and continues until the last beta-strand of the pZBD fold. The overall secondary structure and compactness of wild-type Ec-GluRS remains unaltered in the chimeric constructs. The association of GluRS with tRNAGlu but not with ATP is sensitive to pZBD perturbations. Except for Ec(Bt)-GluRS, all pZBD-chimeras show 100fold or more reduced catalytic efficiency and contain zinc. Natively zinc-bound Ec-GluRS does not require zinc to be active
additional information
-
generation of pZBD-chimeras of Ec-GluRS, four chimeric versions with partly replaced zinc-binding motif, pZBD. In the first chimera [Ec(Te)-GluRS], the pZBD of Ec-GluRS is replaced by the corresponding pZBD from Thermosynechococcus elongatus GluRS (Te-GluRS) whose structure is devoid of a bound Zn2+ despite containing the modified ZB-motif CxCxnYx3H. In the second chimera [Ec(EQRS)-GluRS] the pZBD of Ec-GluRS is replaced by the corresponding pZBD from Escherichia coli Glu-QRS (Ec-EQRS) whose zinc-bound structure contains the ZB-motif CxCxnYx3C. In the third chimera [Ec(Bt)-GluRS], the pZBD of Ec-GluRS is replaced by a 19-residue stretch from the pZBD of Bt-GluRS which contains a disrupted zinc binding motif CxMx20Yx3W and whose structure is devoid of a bound Zn2+ ion. The 19-residue stretch starts from the residue preceding the fourth zinc-co-ordinating cysteine residue in Ec-GluRS and continues until the last beta-strand of the pZBD fold. The overall secondary structure and compactness of wild-type Ec-GluRS remains unaltered in the chimeric constructs. The association of GluRS with tRNAGlu but not with ATP is sensitive to pZBD perturbations. Except for Ec(Bt)-GluRS, all pZBD-chimeras show 100fold or more reduced catalytic efficiency and contain zinc. Natively zinc-bound Ec-GluRS does not require zinc to be active
additional information
-
construction and overexpression of tRNAGlu variants, the enzyme shows altered activity with the tRNA mutants, structural characterization
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. In contrast to the isolated GluRS catalytic domain, 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
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50
-
50% loss of activity, when temperature is increased gradually at the rate of 1 C per min, without addition of substrate
60
-
complete loss of activity, when temperature is increased gradually at the rate of 1°C per min, without addition of substrate
additional information
additional information
-
tRNAGlu in presence of Mg2+ protects against heat inactivation, Mg2+ alone is much less effective
additional information
-
tRNAGlu and ATP protect efficiently against thermal inactivation, glutamate does not. Weak synergism between ATP and tRNAGlu, no synergism between ATP and glutamate. Highest stabilization with ATP, glutamate and tRNAGlu
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
tRNAGlu in presence of Mg2+ protects against heat inactivation, Mg2+ alone is much less effective
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C or -70°C, 20 mM sodium Hepes, pH 7.2, 0.1 mM EDTA, 0.5 mM DTT, 65% glycerol, stable for at least 1 year
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged chimeric mutant enzyme or catalytic domain of GluRS from strain BL21(DE3) or the temperature sensitive strain JP1449(DE3) by nickel affinity chromatography
-
recombinant His-tagged ERS from strain BL21(DE3) by nickel affinity chromatography
-
recombinant N-terminally His6-SUMO2-Gly-tagged K236E/E328A mutant enzyme from Escherichia coli strain Rosetta2 (DE3) by nickel affinity chromatography and dialysis, tag cleavage by SENP2 protease and another step of nickel affinity chromatography to remove the tag, to over 95% purity, followed by ultrafiltration
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene gluS, recombinant overexpression of C-terminally His-tagged GluRS in Escherichia coli
expression of the His-tagged chimeric mutant enzyme and of the catalytic domain of GluRS in Escherichia coli strain BL21(DE3) or the temperature sensitive strain JP1449(DE3)
-
recombinant expression of enzyme mutant K236E/E328A as N-terminally His6-SUMO2-Gly-tagged enzyme in Escherichia coli strain Rosetta2 (DE3) from plasmid DNA PLQ7619
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
construction of an anticodon-binding domain truncated GluRS catalytic domain and a chimeric protein, constructed from the catalytic domain of Escherichia coli GluRS and the anticodon-binding domain of glutaminyl-tRNA synthetase GlnRS. Both proteins discriminate against tRNAGln. In addition to the anticodon-binding domain, tRNAGln discriminatory elements may be present in the catalytic domain in Escherichia coli GluRS as well
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
drug development
inhibitor glutamyl-sulfamoyl-adenosine, Glu-AMS, derivatives with bactericidal properties and low toxicity for humans can be developed
drug development
-
design of better inhibitors specific for bacterial GluRSs, which are promising targets for antimicrobial therapy
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Dorion, C.; Chenvert, R.; Lacoste, L.; Lapointe, J.
Synthesis of an inhibitor of glutamyl-tRNA synthetase
Bioorg. Med. Chem. Lett.
3
2699-2702
1993
Escherichia coli
-
Lin, S.X.; Brisson, A.; Liu, J.; Roy, P.H.; Lapointe, J.
Higher specific activity of the Escherichia coli glutamyl-tRNA synthetase purified to homogeneity by a six-hour procedure
Protein Expr. Purif.
3
71-74
1992
Escherichia coli
Liu, J.; Gagnon, Y.; Gauthier, J.; Furenlid, L.; L'Heureux, P.J.; Auger, M.; Nureki, O.; Yokoyama, S.; Lapointe, J.
The zinc-binding site of Escherichia coli glutamyl-tRNA synthetase is located in the acceptor-binding domain
J. Biol. Chem.
270
15162-15169
1995
Escherichia coli
Liu, J.; Lin, S.X.; Blochet, J.E.; Pezolet, M.; Lapointe, J.
The glutamyl-tRNA synthetase of Escherichia coli contains one atom of zinc essential for its native conformation and its catalytic activity
Biochemistry
32
11390-11396
1993
Bacillus subtilis, Escherichia coli, Thermus thermophilus
Lapointe, J.; Sll, D.
Glutamyl transfer ribonucleic acid synthetase of Escherichia coli. Purification and propertiers
J. Biol. Chem.
247
4966-4974
1972
Escherichia coli
Lapointe, J.; Sll, D.
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Powers, D.M.; Ginsburg, A.
Monomeric structure of glutamyl-tRNA synthetase in Escherichia coli
Arch. Biochem. Biophys.
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Escherichia coli
Kern, D.; Potier, S.; Boulanger, Y.; Lapointe, J.
The Monomeric glutamyl-tRNA synthetase of Escherichia coli. Purification and relation between its structural and catalytic properties
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Escherichia coli
Kern, D.; Lapointe, J.
The glutamyl-tRNA synthetase of Escherichia coli: substrate-induced protection against its thermal inactivation
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Escherichia coli
Kern, D.; Lapointe, J.
Glutamyl transfer ribonucleic acid synthetase of Escherichia coli. Effect of alteration of the 5-(methylaminomethyl)-2-thiouridine in the anticodon of glutamic acid transfer ribonucleic acid on the catalytic mechanism
Biochemistry
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Escherichia coli
Kern, D.; Lapointe, J.
The catalytic mechanism of the glutamyl-tRNA synthetase from Escherichia coli. Detection of an intermediate complex in which glutamate is activated
J. Biol. Chem.
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Escherichia coli
Kern, D.; Lapointe, J.
The catalytic mechanism of glutamyl-tRNA synthetase of Escherichia coli. A steady-state kinetic investigation
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Escherichia coli
Lapointe, J.; Levasseur, S.; Kern, D.
Glutamyl-tRNA synthetase from Escherichia coli
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Escherichia coli
Bothra, A.K.; Roy, S.; Mandal, C.; Mukhophadhyay, C.
Role of zinc in tRNA-acceptor stem binding by glutamyl-tRNA synthetase from E. coli: a molecular modeling study
J. Biomol. Struct. Dyn.
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1997
Escherichia coli
Desjardins, M.; Garneau, S.; Desgagnes, J.; Lacoste, L.; Yang, F.; Lapointe, J.; Chenevert, R.
Glutamyl adenylate analogs are inhibitors of glutamyl-tRNA synthetase
Bioorg. Chem.
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Escherichia coli
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Sekine, S.I.; Nureki, O.; Tateno, M.; Yokoyama, S.
The identity determinants required for the discrimination between tRNAGlu and tRNAAsp by glutamyl-tRNA synthetase from Escherichia coli
Eur. J. Biochem.
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Escherichia coli
Madore, E.; Florentz, C.; Giege, R.; Sekine, S.; Yokoyama, S.; Lapointe, J.
Effect of modified nucleotides on Escherichia coli tRNAGlu structure and on its aminoacylation by glutamyl-tRNA synthetase. Predominant and distinct roles of the mnm5 and s2 modifications of U34
Eur. J. Biochem.
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Escherichia coli
Bernier, S.; Dubois, D.Y.; Habegger-Polomat, C.; Gagnon, L.P.; Lapointe, J.; Chenevert, R.
Glutamylsulfamoyladenosine and pyroglutamylsulfamoyladenosine are competitive inhibitors of E. coli glutamyl-tRNA synthetase
J. Enzyme Inhib. Med. Chem.
20
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2005
Escherichia coli, Mus musculus
Blaise, M.; Becker, H.D.; Keith, G.; Cambillau, C.; Lapointe, J.; Giege, R.; Kern, D.
A minimalist glutamyl-tRNA synthetase dedicated to aminoacylation of the tRNAAsp QUC anticodon
Nucleic Acids Res.
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Escherichia coli
Dubois, D.Y.; Blaise, M.; Becker, H.D.; Campanacci, V.; Keith, G.; Giege, R.; Cambillau, C.; Lapointe, J.; Kern, D.
An aminoacyl-tRNA synthetase-like protein encoded by the Escherichia coli yadB gene glutamylates specifically tRNAAsp
Proc. Natl. Acad. Sci. USA
101
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2004
Escherichia coli
Salazar, J.C.; Ambrogelly, A.; Crain, P.F.; McCloskey, J.A.; Soll, D.
A truncated aminoacyl-tRNA synthetase modifies RNA
Proc. Natl. Acad. Sci. USA
101
7536-7541
2004
Escherichia coli
Balg, C.; Blais, S.P.; Bernier, S.; Huot, J.L.; Couture, M.; Lapointe, J.; Chenevert, R.
Synthesis of beta-ketophosphonate analogs of glutamyl and glutaminyl adenylate, and selective inhibition of the corresponding bacterial aminoacyl-tRNA synthetases
Bioorg. Med. Chem.
15
295-304
2007
Escherichia coli
Gustilo, E.M.; Dubois, D.Y.; Lapointe, J.; Agris, P.F.
E. coli glutamyl-tRNA synthetase is inhibited by anticodon stem-Loop domains and a minihelix
RNA Biol.
4
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2007
Escherichia coli
Saha, R.; Dasgupta, S.; Basu, G.; Roy, S.
A chimeric glutamyl: glutaminyl-tRNA synthetase: implications for evolution
Biochem. J.
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Escherichia coli
Saha, R.; Dasgupta, S.; Basu, G.; Roy, S.
A chimaeric glutamyl:glutaminyl-tRNA synthetase: Implications for evolution
Biochem. J.
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2009
Escherichia coli (P04805), Escherichia coli
Dasgupta, S.; Saha, R.; Dey, C.; Banerjee, R.; Roy, S.; Basu, G.
The role of the catalytic domain of E. coli GluRS in tRNAGln discrimination
FEBS Lett.
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2009
Escherichia coli
Chongdar, N.; Dasgupta, S.; Datta, A.B.; Basu, G.
Preliminary X-ray crystallographic analysis of an engineered glutamyl-tRNA synthetase from Escherichia coli
Acta Crystallogr. Sect. F
70
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2014
Escherichia coli
Chongdar, N.; Dasgupta, S.; Datta, A.B.; Basu, G.
Dispensability of zinc and the putative zinc-binding domain in bacterial glutamyl-tRNA synthetase
Biosci. Rep.
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2015
Escherichia coli (P04805), Escherichia coli
Blais, S.P.; Kornblatt, J.A.; Barbeau, X.; Bonnaure, G.; Laguee, P.; Chenevert, R.; Lapointe, J.
tRNAGlu increases the affinity of glutamyl-tRNA synthetase for its inhibitor glutamyl-sulfamoyl-adenosine, an analogue of the aminoacylation reaction intermediate glutamyl-AMP mechanistic and evolutionary implications
PLoS ONE
10
e0121043
2015
Escherichia coli (P04805), Escherichia coli
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