Information on EC 6.1.1.17 - Glutamate-tRNA ligase

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

EC NUMBER
COMMENTARY
6.1.1.17
-
RECOMMENDED NAME
GeneOntology No.
Glutamate-tRNA ligase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ATP + L-glutamate + tRNAGlu = AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
concerted mechanism
-
ATP + L-glutamate + tRNAGlu = AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
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
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu = AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
catalytic site structure, substrate binding and reaction mechanism
-
ATP + L-glutamate + tRNAGlu = AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
the major recognition element of the tRNAGlu is U at position 34
-
ATP + L-glutamate + tRNAGlu = AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
random binding of ATP and L-glutamate to the enzyme-tRNA complex
-, P0A636
ATP + L-glutamate + tRNAGlu = AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
the transfer of amino acid to tRNA is accompanied by the protonation of AMP to H-AMP. Subsequent migration of proton to water reduces the stability of the complex and loosens the interface both in the presence and in the absence of AMP. The subsequent undocking of AMP or tRNA then proceeds along thermodynamically competitive pathways. Release of the tRNA acceptor stem is further accelerated by the deprotonation of the alpha-ammonium group on the charging amino acid. The proposed general base is Glu41
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Aminoacylation
-
-
-
-
Aminoacylation
-
-
Aminoacylation
O82462
-
Aminoacylation
-
-
Aminoacylation
-
-
Aminoacylation
-
-
Aminoacylation
Bacillus subtilis 168T, Helicobacter pylori 26695
-
-
-
esterification
-
-
-
-
esterification
-
-
esterification
O82462
-
esterification
-
-
esterification
-
-
esterification
-
-
esterification
Bacillus subtilis 168T, Helicobacter pylori 26695
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Aminoacyl-tRNA biosynthesis
-
Biosynthesis of secondary metabolites
-
Metabolic pathways
-
Porphyrin and chlorophyll metabolism
-
tetrapyrrole biosynthesis I (from glutamate)
-
tRNA charging
-
SYSTEMATIC NAME
IUBMB Comments
L-Glutamate:tRNAGlu ligase (AMP-forming)
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D-GluRS
Thermotoga maritima TM1351
-
-
-
discriminating GluRS
-
-
discriminating GluRS
Thermotoga maritima TM1351
-
-
-
EPRS
-
-
GluRS
-
-
-
-
GluRS
Thermotoga maritima TM1351
-
-
-
GluRS1
Helicobacter pylori GluRS1
-
;
-
GluRS1
Helicobacter pylori GluRS2
-
-
-
GluRSAt
O82462
-
Glutamate--tRNA ligase
-
-
-
-
Glutamate--tRNA ligase
-
-
Glutamate-tRNA synthetase
-
-
-
-
Glutamate-tRNA synthetase
-
-
Glutamic acid translase
-
-
-
-
Glutamic acid translase
-
-
Glutamic acid tRNA ligase
-
-
-
-
Glutamic acid tRNA ligase
-
-
Glutaminyl-tRNA synthetase
-
-
Glutamyl tRNA synthetase
-
-
-
-
Glutamyl tRNA synthetase
-
-
glutamyl-prolyl tRNA synthetase
-
-
glutamyl-Q tRNAASp synthetase
-
-
Glutamyl-transfer ribonucleate synthetase
-
-
-
-
Glutamyl-transfer ribonucleate synthetase
-
-
Glutamyl-transfer ribonucleic acid synthetase
-
-
-
-
Glutamyl-transfer ribonucleic acid synthetase
-
-
Glutamyl-transfer RNA synthetase
-
-
-
-
Glutamyl-transfer RNA synthetase
-
-
Glutamyl-tRNA synthetase
-
-
-
-
Glutamyl-tRNA synthetase
-
-
Glutamyl-tRNA synthetase
-
-
Glutamyl-tRNA synthetase
O82462
-
Glutamyl-tRNA synthetase
-
-
Glutamyl-tRNA synthetase
-
-
Glutamyl-tRNA synthetase
-
-
Glutamyl-tRNA synthetase
-
-
-
Glutamyl-tRNA synthetase
Methanocaldococcus jannaschii, Methanococcus thermoautotrophicum, Methanosarcina mazei
-
-
Glutamyl-tRNA synthetase
-
-
Glutamyl-tRNA synthetase
-
-
Glutamyl-tRNA synthetase
-
-
Glutamyl-tRNA synthetase
P46655
-
Glutamyl-tRNA synthetase
-
-
Glutamyl-tRNA synthetase
-
-
Glutamyl-tRNA synthetase
Tolypothrix sp.
-
-
Glutamyl-tRNA synthetase
-
-
P85
-
-
-
-
surface-exposed glutamyl tRNA synthetase
-
-
TM1351
Thermotoga maritima TM1351
-
-
-
tRNA modifying enzyme
-
-
additional information
-
cf. EC 6.1.1.15
CAS REGISTRY NUMBER
COMMENTARY
9068-76-2
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
; strain ATCC 23270, isoform 1 and 2, genes gltX1 and gltX2
-
-
Manually annotated by BRENDA team
strain ATCC 23270, two isozymes GluRS1 and GluRS2
-
-
Manually annotated by BRENDA team
strain 168 Trp-
-
-
Manually annotated by BRENDA team
strain 168T
-
-
Manually annotated by BRENDA team
Bacillus subtilis 168
strain 168 Trp-
-
-
Manually annotated by BRENDA team
Bacillus subtilis 168T
strain 168T
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Caesalpinia bondue
-
-
-
Manually annotated by BRENDA team
non-discriminating enzyme form
-
-
Manually annotated by BRENDA team
mutants encoding GluRS variants altered in the 98C-138C segment
-
-
Manually annotated by BRENDA team
overproducing strain DH5alpha(pLQ7612)
-
-
Manually annotated by BRENDA team
pure enzyme
-
-
Manually annotated by BRENDA team
pure recombinant enzyme from overproducing strain DH5alpha
-
-
Manually annotated by BRENDA team
strain MRE-600 and overproducing strain HS7611
-
-
Manually annotated by BRENDA team
the YadB protein displays 34% identity with the catalytic core of glutamy-tRNA synthetase but lacks the anticodon-binding domain
-
-
Manually annotated by BRENDA team
yadB gene encodes a truncated GluRS that lacks the C-terminal third of the protein and, consequently the anticodon binding domain
-
-
Manually annotated by BRENDA team
Geobacillus stearothermophilus 168T
strain 168T
-
-
Manually annotated by BRENDA team
isoform GluRS2
-
-
Manually annotated by BRENDA team
strain 26695, isozyme 2
-
-
Manually annotated by BRENDA team
strain 26695, isozyme 2
-
-
Manually annotated by BRENDA team
Helicobacter pylori GluRS1
GluRS1
-
-
Manually annotated by BRENDA team
Methanococcus thermoautotrophicum
-
-
-
Manually annotated by BRENDA team
enzyme has dual substrate specificity for L-glutamate and L-proline, enzyme is part of a large aminoacyl-tRNA synthetases complex
-
-
Manually annotated by BRENDA team
strain RS453
-
-
Manually annotated by BRENDA team
Saccharomyces cerevisiae RS453
strain RS453
-
-
Manually annotated by BRENDA team
yellow pigment mutant C-2A'
-
-
Manually annotated by BRENDA team
no glutaminyl-tRNA synthetase activity found in crude extract, but a specific aminotransferase activity that changes Glu-tRNAGln to Gln-tRNAGln
-
-
Manually annotated by BRENDA team
several strains and cultivars, overview, gene gltX
-
-
Manually annotated by BRENDA team
Thermotoga maritima TM1351
-
-
-
Manually annotated by BRENDA team
discriminating enzyme form, purified recombinant enzyme from overexpressing Escherichia coli strain JM109(DE3)
Uniprot
Manually annotated by BRENDA team
overexpressed in Escherichia coli; strain HB8; wild-type and mutant enzymes
-
-
Manually annotated by BRENDA team
strain HB8; wild-type and mutant enzymes
-
-
Manually annotated by BRENDA team
wild-type and mutant enzymes
-
-
Manually annotated by BRENDA team
Tolypothrix sp.
strain PCC 7601, gene gltX
-
-
Manually annotated by BRENDA team
enzyme forms: GuRSP and GluRSE
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
cytoplasmic glutamyl tRNA synthetase gene ers1 is an Moc3 interacting element. Cell growth is moderately affected by cGluRS over-expression under de-repressed conditions. Over-expression of ers1 stimulates sexual differentiation; mitochondrial glutamyl tRNA synthetase gene ers2 is an Moc3 interacting element. Cell growth is severely affected by ers2 over-expression under de-repressed conditions. Under repressed conditions, the cells multiply quickly. In addition, cells over-expressing ers2 show higher mating efficiency than control
physiological function
-
in cells containing glutaminyl-tRNA synthetase, GlnRS, discriminating GluRS specifically aminoacylates tRNAGlu with glutamate. One of two GluRSs from the extremophile Acidithiobacillus ferrooxidans, is inactivated when intracellular heme is elevated suggesting a specific role for GluRS1 in the regulation of tetrapyrrole biosynthesis
metabolism
-
the sensitivity to oxidation of GluRS1 might provide a means to regulate tetrapyrrole and protein biosynthesis in response to extreme changes in both the redox and heme status of the cell via a single enzyme. The glutamate moiety of Glu-tRNAGlu is transformed to glutamate semialdehyde by the glutamyl-tRNA reductase and is subsequently transformed to 4-aminolevulinic acid, the universal precursor of tetrapyrroles, by the glutamate semialdehyde amidotransferase
additional information
-
targets for oxidation-based inhibition are cysteines from a SWIM zinc-binding motif located in the tRNA acceptor helix-binding domain. Oxidation of the metal-binding site cysteine of GluRS1 significantly impaired catalysis. Also, binding of ATP or tRNA protects the distant cysteines of the SWIM motif
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + Glu + tRNAAsp
AMP + diphosphate + L-glutamyl-tRNAAsp
show the reaction diagram
-
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
show the reaction diagram
-
tRNAGlu is not used as substrate
-
-
?
ATP + Glu + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
-
-
-
?
ATP + Glu + tRNAGlU
AMP + diphosphate + L-glutamyl-tRNAGlU
show the reaction diagram
-
GluRS1
-
-
?
ATP + Glu + tRNAGlU
AMP + diphosphate + L-glutamyl-tRNAGlU
show the reaction diagram
Helicobacter pylori, Helicobacter pylori GluRS1
-
GluRS1 cannot form Glu-tRNAGln
-
-
?
ATP + Glu + tRNAGlU
AMP + diphosphate + L-glutamyl-tRNAGlU
show the reaction diagram
Helicobacter pylori GluRS1, Helicobacter pylori GluRS2
-
GluRS1
-
-
?
ATP + glutamate + chloroplastic tRNAGln
AMP + diphosphate + gluamyl-tRNAGln
show the reaction diagram
-
misacylation
-
-
-
ATP + glutamate + chloroplastic tRNAGln
AMP + diphosphate + gluamyl-tRNAGln
show the reaction diagram
-
misacylation
-
-
-
ATP + L-glutamate + tRNAAsp
AMP + diphosphate + L-glutamyl-tRNAAsp
show the reaction diagram
-
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 + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
enzyme expressed in an Escherichia coli mutant strain, tRNAGln UUG from Escherichia coli
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
enzyme is active with the tRNAGln from Bacillus subtilis and the isoacceptor tRNAGln1, but not tRNAGln2, from Escherichia coli, the major recognition element is U at position 34
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
isozyme 2 expressed in an Escherichia coli mutant strain, tRNAGln UUG from Escherichia coli
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
mitochondrial glutamyl-tRNA synthetase efficiently aminoacylates both tRNAGln to form Glu-tRNAGln and tRNAGlu to form Glu-tRNAGlu
-
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
enzyme expressed in an Escherichia coli mutant strain, tRNAGln UUG from Escherichia coli
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
Caesalpinia bondue
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
P27000
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
P27000, -
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
P04805
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
r
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
r
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-, P0A636
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-, O82462
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
Methanococcus thermoautotrophicum
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
P46655
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
glutamylate E. coli tRNAGluF, not cytoplasmic tRNAGlu from yeast and barley
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
chemically modified tRNAGlu modified by monobromobimane or CNBr is a poor substrate. tRNAGlu from the chloroplast of barley, Chlamydomonas reinhardtii, tobacco, cucumber, wheat, and spinach, and tRNAGlu from Synechocystis PCC6803, Escherichia coli, barley germ and bakers yeast are effective substrates, G10, A26, U35 and A37 are recognition elements of barley enzyme
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
existence of an enzyme-AMP-Glu intermediate in the aminoacylation reaction
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
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
show the reaction diagram
-
in absence of tRNAGlu, GluRS binds to D-glutamate as well as L-glutamate
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
charges tRNAGlu from barley and from E. coli
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
enzyme has dual substrate specificity for L-glutamate and L-proline
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
enzyme utilizes tRNAGlu from Bacillus subtilis, not from Escherichia coli
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
isozyme 1 expressed in an Escherichia coli mutant strain, tRNAGlu from Escherichia coli
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
recombinant mutant Q373R expressed in Escherichia coli mutant strain, tRNAGlu from Escherichia coli
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
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
show the reaction diagram
P27000
tRNAGlu binding causes conformational changes in the enzyme, glutamine binding mechanism, in presence or absence of tRNA
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
wild-type tRNAGlu and tRNA AE(GU)
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
eukaryotic-type discriminating glutamyl-tRNA synthetase, inability to utilize Escherichia coli tRNA as substrate. The enzyme is essential for growth of insect stage Trypanosoma brucei and is responsible for essentially all of the glutamyl-tRNA synthetase activity in cytosol and in mitochondria, eukaryotic-type discriminating glutamyl-tRNA synthetase, inability to utilize Escherichia coli tRNA as substrate
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
glutamyl-tRNA, formed by Glu-tRNA synthetase, is a substrate for protein biosynthesis and tetrapyrrole formation by the C5 pathway
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
a two-step reaction
-
-
r
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
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
show the reaction diagram
-
structural bases of transfer RNA-dependent L-glutamate recognition and activation by the enzyme, the glutamate-binding site is immature in the absence of tRNA, overview
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
wild-type enzyme and chimeric mutant cGluGlnRS, overview, analysis of domain functions in enzyme-substrate interactions, overview
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
mitochondrial glutamyl-tRNA synthetase efficiently aminoacylates both tRNAGln to form Glu-tRNAGln and tRNAGlu to form Glu-tRNAGlu
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
Bacillus subtilis 168
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
Bacillus subtilis 168
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
Saccharomyces cerevisiae RS453
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
Geobacillus stearothermophilus 168T
-
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
Bacillus subtilis 168T
-
-
-
?
ATP + L-glutamate + tRNAGlu
?
show the reaction diagram
-
involved in synthesis of 5-aminolevulinate (a committed and regulated precursor in the chlorophyll biosynthetic pathway)
-
-
-
ATP + L-glutamate + tRNAGlu
?
show the reaction diagram
-
involved in synthesis of 5-aminolevulinate (a committed and regulated precursor in the chlorophyll biosynthetic pathway)
-
-
-
ATP + L-glutamate + tRNAGlu mutant C36G
AMP + diphosphate + L-glutamyl-tRNAGlu mutant C36G
show the reaction diagram
P27000, -
mutant R358Q, low activity with the wild-type enzyme
-
?
ATP + L-glutamate + tRNAGlu wild-type
AMP + diphosphate + L-glutamyl-tRNAGlu wild-type
show the reaction diagram
P27000, -
enzyme is specific for tRNAGlu
-
?
ATP + L-proline + tRNAGlu
AMP + diphosphate + L-prolyl-tRNAGlu
show the reaction diagram
-
enzyme has dual substrate specificity for L-glutamate and L-proline
-
?
additional information
?
-
-
ATP-diphosphate exchange, threo-4-methyl-DL-glutamic acid or threo-4-hydroxy-L-glutamic acid can promote ATP-diphosphate exchange
-
-
-
additional information
?
-
Caesalpinia bondue
-
threo-4-methyl-DL-glutamic acid or threo-4-hydroxy-L-glutamic acid can promote ATP-diphosphate exchange
-
-
-
additional information
?
-
-
erythro-4-methyl-L-glutamic acid, erythro-4-hydroxy-DL-glutamic acid, or 2(S),4(S)-4-hydroxy-4-methyl-L-glutamic acid can promote ATP-diphosphate exchange
-
-
-
additional information
?
-
-
tRNAGlu-dependent ATP-diphosphate exchange
-
-
-
additional information
?
-
-
glutamate and glutamine acceptor activity with wild-type and mutant tRNAGlns with native and recombinant enzyme, overview
-
?
additional information
?
-
-
no charging of Escherichia coli tRNAGln by enzyme mutant Q373R
-
?
additional information
?
-
-
no recognition of recombinant tRNA mutants AQ(GU) and AQ(GC)
-
?
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 recombinant wild-type enzyme is toxic for Escherichia coli, probably due to its charging of both tRNAGlu and tRNAGln
-
?
additional information
?
-
-
the enzyme also catalyzes ATP-diphosphate exchange
-
-
-
additional information
?
-
-
GluRS plays a major role in regulating the cellular level of heme, aminoacylation of tRNAGlu correlates with the demand of heme, a transcriptional mechanism might control the level of GluRS1 in cells grown in Fe2+, under growth conditions in which cells do not require Glu-tRNA, as precursor for heme biosynthesis, up to 85% of GluRS1 is dispensable, but no major detrimental effect in the cell growth is observed. Thus, GluRS2 and the remaining 15% of the activity of GluRS1 are sufficient to provide the Glu-tRNA substrates for protein synthesis
-
-
-
additional information
?
-
-
GtS is an age-dependent Streptococcus pneumoniae antigen and is a surface-located adhesin that is capable of inducing a partially protective immune response against Streptococcus pneumoniae in mice, overview
-
-
-
additional information
?
-
Tolypothrix sp.
-
regulation of gltX expression, overview, the gene glxT encoding the enzyme is involved in regulation of other genes's expression, mechanisms, overview
-
-
-
additional information
?
-
P46655
GluRS interacts with the accessory protein Arc1p, interaction mode and structure, overview
-
-
-
additional information
?
-
-
GluRS interacts with the accessory protein Arc1p, interaction mode and structure, overview
-
-
-
additional information
?
-
-
substrate and co-factor recognition and binding structures, GluRS and tRNAGlu collaborate to form a highly complementary L-glutamate-binding site, the collaborative site is functional, amino acid specificity is generated in the GluRS-tRNA complex, overview
-
-
-
additional information
?
-
-
thermal stability and structural analysis of tRNA substrates, overview
-
-
-
additional information
?
-
-
the glutamyl-prolyl tRNA synthetase determines the specificity of the heterotetrameric GAIT complex suppressing translation of selected mRNAs in interferon-gamma-activated monocytic cells by binding to a 3' UTR element in target mRNAs, critical role of EPRS WHEP domains in targeting and regulating GAIT complex binding to RNA, mechanism, overview. The enzyme is essential in regulating inflammatory gene expression, the upstream WHEP pair of EPRS directs high-affinity binding to GAIT element-bearing mRNAs, while the overlapping, downstream pair binds NSAP1, which inhibits mRNA binding. Interaction of EPRS with ribosomal protein L13a and GAPDH induces a conformational witch that rescues mRNA binding and restores translational control, interaction analysis, overview
-
-
-
additional information
?
-
-
isoform GluRS2 is unable to produce Glu-tRNAGlu. Within a series of tRNA chimeras containing 75% tRNAGln and 25% tRNAGlu2 character, GluRS2 recognizes major identity elements clustered in the tRNAGln acceptor stem. Mutations in the tRNA anticodon or at the discriminator base have little to no impact on enzyme specificity and activity
-
-
-
additional information
?
-
-
D-GluRS glutamylates tRNAGlu only
-
-
-
additional information
?
-
-
discriminating GluRS specifically aminoacylates tRNAGlu with glutamate. Acidithiobacillus ferrooxidans GluRS1 contains cysteines 98, 100 and 125 together with glutamate 127 clustered in the catalytic domain
-
-
-
additional information
?
-
Thermotoga maritima TM1351
-
D-GluRS glutamylates tRNAGlu only
-
-
-
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 + L-glutamyl-tRNAGln
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
enzyme expressed in an Escherichia coli mutant strain, tRNAGln UUG from Escherichia coli
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
isozyme 2 expressed in an Escherichia coli mutant strain, tRNAGln UUG from Escherichia coli
-
?
ATP + L-glutamate + tRNAGln
AMP + diphosphate + L-glutamyl-tRNAGln
show the reaction diagram
-
enzyme expressed in an Escherichia coli mutant strain, tRNAGln UUG from Escherichia coli
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
P27000
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
P27000, -
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
r
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
r
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-, O82462
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
Methanococcus thermoautotrophicum
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
P46655
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
isozyme 1 expressed in an Escherichia coli mutant strain, tRNAGlu from Escherichia coli
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
recombinant mutant Q373R expressed in Escherichia coli mutant strain, tRNAGlu from Escherichia coli
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
eukaryotic-type discriminating glutamyl-tRNA synthetase, inability to utilize Escherichia coli tRNA as substrate. The enzyme is essential for growth of insect stage Trypanosoma brucei and is responsible for essentially all of the glutamyl-tRNA synthetase activity in cytosol and in mitochondria
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
glutamyl-tRNA, formed by Glu-tRNA synthetase, is a substrate for protein biosynthesis and tetrapyrrole formation by the C5 pathway
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
wild-type enzyme and chimeric mutant cGluGlnRS, overview
-
-
?
ATP + L-glutamate + tRNAGlu
?
show the reaction diagram
-
involved in synthesis of 5-aminolevulinate (a committed and regulated precursor in the chlorophyll biosynthetic pathway)
-
-
-
ATP + L-glutamate + tRNAGlu
?
show the reaction diagram
-
involved in synthesis of 5-aminolevulinate (a committed and regulated precursor in the chlorophyll biosynthetic pathway)
-
-
-
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
-
-
-
?
ATP + L-glutamate + tRNAGlu
AMP + diphosphate + L-glutamyl-tRNAGlu
show the reaction diagram
Bacillus subtilis 168T
-
-
-
?
additional information
?
-
-
no charging of Escherichia coli tRNAGln by enzyme mutant Q373R
-
?
additional information
?
-
-
the recombinant wild-type enzyme is toxic for Escherichia coli, probably due to its charging of both tRNAGlu and tRNAGln
-
?
additional information
?
-
-
GluRS plays a major role in regulating the cellular level of heme, aminoacylation of tRNAGlu correlates with the demand of heme, a transcriptional mechanism might control the level of GluRS1 in cells grown in Fe2+, under growth conditions in which cells do not require Glu-tRNA, as precursor for heme biosynthesis, up to 85% of GluRS1 is dispensable, but no major detrimental effect in the cell growth is observed. Thus, GluRS2 and the remaining 15% of the activity of GluRS1 are sufficient to provide the Glu-tRNA substrates for protein synthesis
-
-
-
additional information
?
-
-
GtS is an age-dependent Streptococcus pneumoniae antigen and is a surface-located adhesin that is capable of inducing a partially protective immune response against Streptococcus pneumoniae in mice, overview
-
-
-
additional information
?
-
Tolypothrix sp.
-
regulation of gltX expression, overview, the gene glxT encoding the enzyme is involved in regulation of other genes's expression, mechanisms, overview
-
-
-
additional information
?
-
-
the glutamyl-prolyl tRNA synthetase determines the specificity of the heterotetrameric GAIT complex suppressing translation of selected mRNAs in interferon-gamma-activated monocytic cells by binding to a 3' UTR element in target mRNAs, critical role of EPRS WHEP domains in targeting and regulating GAIT complex binding to RNA, mechanism, overview. The enzyme is essential in regulating inflammatory gene expression
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ATP
-
binding mechanism, in presence or absence of tRNA
ATP
-
ATP binds to the 'productive' subsite, due to the tRNA-induced rearrangement of the binding site, which is, at least partially, the structural basis of the tRNA-dependent enzyme activation for amino acid activation
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
Caesalpinia bondue
-
Km: 2.2 mM, ATP-diphosphate exchange, 6.3 mM, glutamate-tRNA formation; required
Mg2+
-
Km: 3.4 mM, ATP-diphosphate exchange; required
Mg2+
-
Km: 1.8 mM, ATP-diphosphate exchange, 4.8 mM, glutamate-tRNA formation; required
Mg2+
-
optimal concentration is 10 mM; required
Mg2+
-
Km: 0.28 mM, enzyme form GluRSE, aminoacylation; Km: 0.46 mM, enzyme form GluRSC, aminoacylation, Km: 0.38 mM, enzyme form GluRSP, aminoacylation; required
Mg2+
-
required
Mg2+
-
required
Mg2+
-
-
Mn2+
-
can partially replace Mg2+ in activation, maximal efficiency at 5 mM is 71% of Mg2+-activation
Zinc
-
zinc metalloenzyme
Zinc
-
enzyme contains one zinc atom strongly bound, which is essential for its native conformation and its catalytic acitivity; zinc metalloenzyme
Zinc
-
enzyme does not contain zinc
Zinc
-
enzyme molecule contains one atom of 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 metalloenzyme
Zn2+
-
bound at the active site, Zn2+ is essential for the proper binding of glutamate to GluRS. GluRS1 contains one Zn2+ ion per enzyme molecule, determination of the Zn2+ content of GluRS1 by mass spectrometry, overview. A relatively canonical SWIM motif, C-X-C-X24-C-X-E, forms a Zn2+-binding site
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(1R,2R)-1-(4-methylsulfonylphenyl)-2-(L-methionylsulfone-amido)-1,3-propadiol
-
-
-
(1R,2R)-1-(4-nitrophenyl)-2-(L-ethionyl-sulfoneamido)-1,3-propadiol
-
competitive inhibition with respect to Asp-tRNAAsn
(1R,2R)-1-phenyl-2-(L-methionyl-sulfone-amido)-1,3-propadiol
-
-
-
(1R,2S)-1-(4-nitrophenyl)-2-(L-methionyl-sulfoneamido)-1,3-propadiol
-
-
(1S,2R)-1-(4-nitrophenyl)-2-(L-methionyl-sulfoneamido)-1,3-propadiol
-
-
(1S,2S)-1-(4-nitrophenyl)-2-(L-methionyl-sulfone-amido)-1,3-propadiol
-
-
1,10-phenanthroline
-
ATP protects the enzyme against zinc removal
5'-O-(N'-(L-pyroglutamyl)-sulfamoyl)adenosine
-
weak
5'-O-(N-(L-glutamyl)-sulfamoyl)adenosine
-
potent competitive with respect to glutamic acid
amino levulinic acid
-
indirect inhibition, growth of Acidithiobacillus ferrooxidans in aminolevulic acid inhibits the activity of GluRS1, the reduced activity of GluRS1 is the result of an interaction of the enzyme with heme or any other intermediate tetrrapyrrole, amino levulic acid added to the reaction mixture has no effect in the activity of GluRSs
Chloramphenicol
-
-
diphosphate
-, P0A636
competitive to ATP and L-glutamate, uncompetitive to tRNAGlu
erythro-4-Hydroxy-DL-glutamic acid
-
glutamate transfer to tRNA
erythro-Methyl-L-glutamic acid
-
glutamate transfer to tRNA
-
gamma-Globulin
-
-
-
glutaminyl-beta-ketophosphonate-adenosine
-
i.e. Gln-KPA, competitive inhibition, non-cognate, binds at one site on the monomeric enzyme
-
glutamol-AMP
-
competitive inhibition
glutamol-AMP
-, P0A636
noncompetitive to ATP and L-glutamate
glutamyl adenylate
-
-
glutamyl cytidylate
-
weak inhibition
glutamyl dihydrocytidylate
-
very weak inhibition
glutamyl N6-benzoyladenylate
-
-
glutamyl uridylate
-
very weak inhibition
glutamyl-beta-ketophosphonate-adenosine
-
i.e. Glu-KPA, selective, competitive inhibition of GluRS, binds at one site on the monomeric enzyme
-
H2O2
-
GluRS1 activity is reversibly inactivated upon oxidation by hydrogen peroxide, the enzyme loses 90% activity after 10 min at 0.3 mM H2O2. tRNAGlu is able to protect GluRS1 against oxidative inactivation by hemin plus hydrogen peroxide. GluRS1 is the main enzyme responsible for supplying Glu-tRNAGlu for heme biosynthesis. Partial recovery of the enzymatic activity by treatment with DTT or 2-mercaptoethanol
heme
-
indirect mechanism, when intracellular heme is in excess, the cells respond by a dramatic decrease of GluRS activity, heme or any other precursor tetrapyrrole is the intracellular effector that triggers this regulatory mechanism
Hemin
-
recombinant GluRS1 enzyme is inhibited in vitro by hemin, but NADPH restores its activity, GluRS2 is also inhibited by hemin to a similar extent as GluRS1
Hemin
-
GluRS1 activity is reversibly inactivated upon oxidation by hemin. tRNAGlu is able to protect GluRS1 against oxidative inactivation by hemin plus hydrogen peroxide
KCl
-
100 mM, 15% inhibition of ATP-diphosphate exchange, 500 mM, 62% inhibition
KCl
-
aminoacylation
N6-Benzoyl-L-glutamyl AMP
-
specific for glutamyl-tRNA synthetase, does not inhibit glutaminyl-tRNA synthetase
NaCl
-
100 mM, 40% inhibition of ATP-diphosphate exchange, 500 mM, 91% inhibition
NH4Cl
-
100 mM, 31% inhibition of ATP-diphosphate exchange, 500 mM, 82% inhibition of ATP-diphosphate exchange
p-hydroxymercuribenzoate
-
-
threo-4-Hydroxy-L-glutamic acid
Caesalpinia bondue, Vigna radiata var. radiata
-
glutamate transfer to tRNA
threo-4-Methyl-D-glutamic acid
Caesalpinia bondue, Vigna radiata var. radiata
-
glutamate transfer to tRNA
LiCl
-
100 mM, 22% inhibition of ATP-diphosphate exchange, 500 mM, 73% inhibition
additional information
-
rGtS and anti-rGtS antiserum significantly inhibits the adhesion of 3 pairs of encapsulated and unencapsulated strains of Streptococcus pneumoniae to human epithelial A549 cells
-
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 ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Arc1p
-
protein from Aspergillus fumigatus, Accession code Q4QTS4, mediates stimulation of tRNA glutamylation
-
Arc1p
-
accessory protein Arc1p, structure of the interacting domains, crystals structure determination and analysis at 1.9 A resolution, phasing, model building, and refinement of Arc1p, unusual noncrystallographic symmetry in the structure of Arc1p, overview
-
Arc1p
-
accessory protein Arc1p, SwissProt ID P46672
-
Arc1p-N
-
residues 1-122 of Arc1p, recombinantly expressed, the GluRS-NArc1p-N complex represents an unusual mode of interaction
-
Bovine serum albumin
-
optimal activity at 0.25 mg/ml
-
tRNA
-
GluRS is one of the aminoacyl-tRNA synthetases that require the cognate tRNA for specific amino acid recognition and activation, tRNA serves as the enzyme activator in the first step, and as the substrate in the second step of aminoacylation, overview, On the other hand, the main chain of the glutamate is immature glutamate-binding site in the absence of tRNA
Cytoplasmic protein Arc1p
-
forms a complex with glutamyl-tRNA synthetase, facilitates the delivery of tRNA molecules to the Arc1p-associated aminoacyl-tRNA synthetase
-
additional information
-
under high heme requirement for respiration increased levels of GluRS occur
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.023
-
ATP
-
wild-type enzyme
0.08
-
ATP
-, P0A636
pH 7.3, 37C
0.1
-
ATP
-
ATP, , 56000 MW subunit and 46000 MW subunit
0.1
-
ATP
-
Glu
0.18
-
ATP
-
-
0.2
-
ATP
-
-
0.25
-
ATP
-
56000 MW subunit alone
0.32
-
ATP
-
ATP-diphosphate exchange
0.043
-
C36G mutant tRNAGlu
P27000
recombinant wild-type enzyme, 65C
-
0.055
-
C36G mutant tRNAGlu
P27000
recombinant mutant R358Q, 65C
-
0.0023
-
Glu
-
-
0.01
-
Glu
-
enzyme form GluRSC, aminoacylation
0.027
-
Glu
-
56000 MW subunit and 46000 MW subunit
0.05
-
Glu
-
-
0.083
-
Glu
-
enzyme forms GluRSP and GluRSE, aminoacylation
0.086
-
Glu
-
56000 MW subunit alone
0.2
-
Glu
-
-
0.07
-
L-Glu
-
-
0.12
-
L-Glu
-
wild-type
0.00042
-
tRNA1Glu
-
-
0.00046
-
tRNA2Glu
-
-
0.00024
-
tRNA3Glu
-
-
-
0.00015
-
tRNAAsp
-
-
0.0017
-
tRNAGln
-
-
0.00008
-
tRNAGlu
-
-
0.0001
-
tRNAGlu
-
-
0.00012
-
tRNAGlu
-
56000 MW subunit alone
0.0002
-
tRNAGlu
-
enzyme forms GluRSC and GluRSP, aminoacylation
0.0002
-
tRNAGlu
-
pH 7.2, 37C, recombinant His-tagged enzyme
0.00027
-
tRNAGlu
-
-
0.0003
-
tRNAGlu
-
56000 MW subunit and 46000 MW subunit
0.00044
-
tRNAGlu
-
enzyme form GluRSE, aminoacylation
0.00045
-
tRNAGlu
-
in presence of the GluRS-Arc1p complex
0.0006
-
tRNAGlu
-
substrate from E. coli
0.00065
-
tRNAGlu
-
substrate from Thermus thermophilus
0.0019
-
tRNAGlu
P04805
wild-type
0.00273
-
tRNAGlu
-
wild-type enzyme
0.056
-
tRNAGlu
-
wild-type tRNA substrate, pH 7.2, 37C
0.7
-
tRNAGlu
-, P0A636
pH 7.3, 37C
0.0047
-
wild-type tRNAGlu
P27000
recombinant wild-type enzyme, 65C
-
0.085
-
wild-type tRNAGlu
P27000
recombinant mutant R358Q, 65C
-
2.7
-
L-glutamate
-, P0A636
pH 7.3, 37C
additional information
-
additional information
-
Km-values of mutant enzymes
-
additional information
-
additional information
-
-
-
additional information
-
additional information
-
-
-
additional information
-
additional information
-
as the KCl concentration is raised from 0 to 100 mM, the Km value for L-glutamate in the reaction with E. coli tRNAGlu is remarkably increased wheras the Km value for glutamate with Thermus thermophilus tRNAGlu is slightly increased
-
additional information
-
additional information
-
Km for diverse modified forms of tRNAAsp and tRNAGlu
-
additional information
-
additional information
-
kinetics, diverse tRNAGlu mutants
-
additional information
-
additional information
-
steady-state kinetics of isozymes GluRS1 and GluRS2
-
additional information
-
additional information
-
kinetics
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.272
-
ATP
-, P0A636
pH 7.3, 37C
0.18
-
C36G mutant tRNAGlu
P27000
recombinant wild-type enzyme, 65C
-
1.4
-
C36G mutant tRNAGlu
P27000
recombinant mutant enzyme R358Q, 65C
-
0.3
-
Glu-tRNA
-
-
-
3.4
-
Glu-tRNA
-
-
-
0.8
-
GlutRNA
-
-
-
0.95
-
tRNAGln
-
-
0.0058
-
tRNAGlu
P04805
wild-type
0.016
-
tRNAGlu
-
in presence of the GluRS-Arc1p complex
0.31
-
tRNAGlu
-
-
0.408
-
tRNAGlu
-, P0A636
pH 7.3, 37C
3
-
tRNAGlu
-
wild-type tRNA substrate, pH 7.2, 37C
1.5
-
wild-type tRNAGlu
P27000
recombinant mutant enzyme R358Q, 65C
-
2.1
-
wild-type tRNAGlu
P27000
recombinant wild-type enzyme, 65C
-
2.15
-
L-glutamate
-, P0A636
pH 7.3, 37C
additional information
-
additional information
-
turnover numbers of wild-type and mutant enzymes
-
additional information
-
additional information
-
activity with diverse modified forms of tRNAAsp and tRNAGlu
-
additional information
-
additional information
-
diverse tRNAGlu mutants
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
560
-
tRNAGln
-
-
17505
1148
-
tRNAGlu
-
-
17506
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.12
-
(1R,2R)-1-(4-methylsulfonylphenyl)-2-(L-methionylsulfone-amido)-1,3-propadiol
-
pH 7.0, temperature not specified in the publication
-
0.027
-
(1R,2R)-1-(4-nitrophenyl)-2-(L-ethionyl-sulfoneamido)-1,3-propadiol
-
pH 7.0, temperature not specified in the publication
0.4
-
(1R,2R)-1-phenyl-2-(L-methionyl-sulfone-amido)-1,3-propadiol
-
pH 7.0, temperature not specified in the publication
-
0.37
-
(1R,2S)-1-(4-nitrophenyl)-2-(L-methionyl-sulfoneamido)-1,3-propadiol
-
pH 7.0, temperature not specified in the publication
2.8
-
(1S,2R)-1-(4-nitrophenyl)-2-(L-methionyl-sulfoneamido)-1,3-propadiol
-
pH 7.0, temperature not specified in the publication
0.16
-
(1S,2S)-1-(4-nitrophenyl)-2-(L-methionyl-sulfone-amido)-1,3-propadiol
-
pH 7.0, temperature not specified in the publication
0.015
-
5'-O-(N'-(L-pyroglutamyl)-sulfamoyl)adenosine
-
-
0.0000028
-
5'-O-(N-(L-glutamyl)-sulfamoyl)adenosine
-
-
0.00007
-
5'-O-(N-(L-glutamyl)-sulfamoyl)adenosine
-
-
1.85
-
Chloramphenicol
-
pH 7.0, temperature not specified in the publication
0.027
-
diphosphate
-, P0A636
substrate ATP, pH 7.3, 37C
0.101
-
diphosphate
-, P0A636
substrate L-glutamate, pH 7.3, 37C
0.0000028
-
Glu-AMS
-
pH 7.2, 37C, versus L-glutamate
2.9
-
glutaminyl-beta-ketophosphonate-adenosine
-
pH 7.2, 37C, versus L-glutamate
-
0.0012
-
glutamol-AMP
-
pH 7.5, 65C
0.0015
-
glutamol-AMP
-, P0A636
substrate ATP, pH 7.3, 37C
0.0039
-
glutamol-AMP
-, P0A636
substrate L-glutamate, pH 7.3, 37C; substrate tRNAGlu, pH 7.3, 37C
0.003
-
glutamyl adenylate
-
37C
0.63
-
glutamyl cytidylate
-
37C
16.7
-
glutamyl dihydrocytidylate
-
37C
0.06
-
glutamyl N6-benzoyladenylate
-
37C
2.75
-
glutamyl uridylate
-
37C
0.018
-
glutamyl-beta-ketophosphonate-adenosine
-
pH 7.2, 37C, versus L-glutamate
-
additional information
-
additional information
-
inhibition kinetics
-
additional information
-
additional information
-
kinetics of inhibitory tRNA fragments, overview
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.0148
-
Caesalpinia bondue
-
-
0.036
-
-
wheat germ enymes
0.0396
-
-
-
0.06
-
-
chloroplastic enzyme
0.064
-
-
-
0.16
-
-
purified recombinant enzyme expressed in Bacillus subtilis
0.79
-
-
strain HRE-600
1.071
-
-
-
1.31
-
-
overproducing strain HS7611
1.95
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
glutamate and glutamine acceptor activity with wild-type and mutant tRNAGlns with native and recombinant enzyme, overview
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.2
-
-
tRNAGlu-dependent ATP-diphosphate exchange
7
-
-
assay at
7
-
-
assay at
7.2
-
-
assay at
7.2
-
-
assay at
7.2
-
-
assay at
7.2
-
-
assay at
7.2
-
-
assay at
7.2
-
-
assay at
7.5
-
-
assay at
7.8
8.2
-
ATP-diphosphate exchange
8
8.5
-
aminoacylation
8
9
-
aminoacylation, in presence of 5 mM Mg2+
8.2
-
-
glutamyl-tRNA formation
8.6
-
-
glutamyl-tRNA formation
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
8.2
-
7: rapid drop of activity below, 7.8-8.2: maximal activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
assay at
37
-
-
12 min assay
37
-
-
assay at
37
-
-
assay at
37
-
-
assay at
37
-
-
assay at
37
-
-
assay at
47
-
-
aminoacylation
65
-
-
aminoacylation
65
-
-
assay at
65
-
P27000
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35
55
-
35C: about 70% of maximal activity, 55C: about 60% of maximal activity
50
80
-
about 25% of maximal activity at 50C and 80C
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
2 enzyme forms: GluRSP and GluRSE
Manually annotated by BRENDA team
Caesalpinia bondue, Vigna radiata var. radiata
-
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Borrelia burgdorferi (strain ATCC 35210 / B31 / CIP 102532 / DSM 4680)
Burkholderia thailandensis (strain E264 / ATCC 700388 / DSM 13276 / CIP 106301)
Methanothermobacter thermautotrophicus (strain ATCC 29096 / DSM 1053 / JCM 10044 / NBRC 100330 / Delta H)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Thermosynechococcus elongatus (strain BP-1)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
58000
-
-
electrophoresis in polyacrylamide gels of various concentrations
60000
-
-
HPLC gel filtration
62000
-
-
sucrose density gradient centrifugation HPLC gel filtration
62000
-
-
gel filtration
102000
-
-
calculation from amino acid composition
105000
-
-
gel filtration
110000
112000
-
non-denaturing PAGE, gel filtration, chloroplastic enzyme
111000
-
-
gel filtration
155000
160000
-
gel filtration, non-denaturing PAGE, enzyme form GluRSP
164000
165000
-
non-denaturing PAGE, gel filtration, enzyme form GluRSE
172000
-
-
-
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 54000, SDS-PAGE
?
-
x * 55671, calculation from nucleotide sequence
?
-
x * 54000, SDS-PAGE
?
-
x * 53901, calculation from nucleotide sequence
?
-
x * 55000, recombinant enzyme expressed in Bacillus subtilis, SDS-PAGE
?
O82462
x * 81000, recombinant protein, SDS-PAGE
?
-
x * 58000, recombinant enzyme, SDS-PAGE
?
Geobacillus stearothermophilus 168T
-
x * 55671, calculation from nucleotide sequence
-
dimer
-
2 * 32500, SDS-PAGE
dimer
-
1 * 56000 (catalytic subunit) + 1 * 46000 (no detectable enzymatic activity, protective function), SDS-PAGE
dimer
-
2 * 55000, SDS-PAGE, chloroplastic enzyme; 2 * 79000, SDS-PAGE, enzyme form GluRSP; 2 * 83000, SDS-PAGE, enzyme form GluRSE
dimer
-
the active forms are dimers, which are in equilibrium with inactive monomers
dimer
-
2 * 55000, SDS-PAGE
monomer
-
1 * 62000, SDS-PAGE
monomer
-
1 * 62000, SDS-PAGE
monomer
-
1 * 56000, SDS-PAGE
monomer
-
1 * 65500, enzyme copurifies with a 46000 MW polypeptide which increases the affinity for glutamate and ATP, and stabilizes it against heat inactivation, SDS-PAGE
monomer
-
1 * 55000, recombinant His6-tagged GluRS, SDS-PAGE
monomer
Bacillus subtilis 168
-
1 * 65500, enzyme copurifies with a 46000 MW polypeptide which increases the affinity for glutamate and ATP, and stabilizes it against heat inactivation, SDS-PAGE
-
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
-
structure and organization of the multienzyme complex
additional information
-
the pretransition-state quaternary complex, crystal structure analysis, in the GluRS-tRNAGlu-Glu structure, GluRS and tRNAGlu collaborate to form a highly complementary L-glutamate-binding site
additional information
-
the human EPRS consists of four major domains: an N-terminal elongation factor-1Bgamma-like domain, an ERS catalytic domain, a 300 amino acid linker domain containing three tandem WHEP domains, and a C-terminal PRS catalytic domain
additional information
-
structure analysis, overview
additional information
-
Acidithiobacillus ferrooxidans GluRS1 contains cysteines 98, 100 and 125 together with glutamate 127 clustered in the catalytic domain
additional information
Thermotoga maritima TM1351
-
structure analysis, overview
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
lipoprotein
-
high molecular weight aminoacyl-tRNA synthetase complex contains lipid. Delipidation does not affect the size or activity of the complex, but a variety of functional and structural properties of individual synthetases in the complex are altered: sensitivity to salts plus detergents, temperature inactivation, hydrophobicity, sensitivity to protease digestion
additional information
-
GluRS is substrate of DsbA, a protein involved in the restoration of the reduced state of cysteines in proteins upon oxidation
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
purified recombinant GluRS-N-Arc1p-N complex, hanging drop vapour diffusion method, 0.002 ml protein solution containing 15 mg/ml protein in 20 mM HEPES, 150 mM NaCl, 5 mM MgCl2, 1 mM DTT, pH 7.2 with NaOH, is mixed with 0.002 ml reservoir solution containing 30-35% PEG 3350, 300-500 mM NaSCN, X-ray diffraction structure determination and analysis at 2.05 A resolution
-
purified recombinant truncated enzyme, 0.002 ml of 20 mg/ml protein in 20 mM HEPES, 150 mM NaCl, 5 mM MgCl2, 1 mM DTT, pH 7.2 with NaOH, mixed with 0.002 ml reservoir solution at 20C, equilibration against 0.075 ml reservoir solution, containing 1.7-1.8 M (NH4)2SO4, 200 mM KSCN for selenomethionine-substituted crystals and 1.8-1.9 M (NH4)2SO4, 200 mM NaI for native crystals, dispersion with selenomethionine, X-ray diffraction structure determination and analysis at 2.5 A resolution, modeling
-
architectures of class-defining and specific domains
-
crystallization of complexes: 1. GluRS and L-Glu, 2. GluRS, tRNAGlu, and L-Glu, 3. GluRS, tRNAGlu, ATP, and L-glutamol, 4. GluRS, tRNAGlu, and L-glutamyl-sulfamoyl adenosine, by hanging drop vapour diffusion method, 5.0 mg/ml enzyme in 10 mM MOPS-Na buffer, pH 6.5, MgCl2, 5 mM 2-mercaptoethanol, 1% PEG 6000, and 2 mM L-glutamate, equilibration against a 1 ml reservoir solution containing 10% PEG at 4C, ERS/tRNA/Glu and ERS/tRNA/ESA crystals are prepared by diffusing 1 mM L-glutamate and 0.5 mM glutamyl-sulfamoyl adenosine, i.e. ESA, respectively, into the ERS/tRNA binary complex crystals, ERS/tRNA/ATP/Eol crystals are obtained by adding both 1 mM ATP and 1 mM L-glutamol, i.e. Eol, to drops containing the ERS/tRNA binary complex, X-ray diffraction structure determination and analysis at 1.98 A, 2.4 A, 2.2 A, and 2.69 A resolution, respectively
-
crystallization of the enzyme in different complexes: 1. non-productively complexed with ATP and L-glutamate, 2. with ATP, 3. with tRNAGlu and ATP, 4. with tRNAGlu and the glutamyl-AMP analogue glutamol-AMP, hanging-drop method, 0.008 ml of 5.0 mg/ml protein in 10 mM Na-MOPS, pH 6.5, 5 mM MgCl2, 2.5 mM 2-mercaptoethanol, 1% PEG 6000, 1-2 mM ATP and/or 2 mM glutamate and/or 0.5 mM glutamol-AMP, plus 1 ml reservoir solution containing 10% PEG 6000 at 4 or 20C, 3 days or more, X-ray diffraction structure determination at 1.8 A resolution, molecular replacement, and analysis
-
molecular modeling, internal pKa calculations, and molecular dynamics simulations for consideration of distinct, mechanistically relevant post-transfer states with charged tRNA bound to glutamyl-tRNA synthetase. The transfer of amino acid to tRNA is accompanied by the protonation of AMP to H-AMP. Subsequent migration of proton to water reduces the stability of the complex and loosens the interface both in the presence and in the absence of AMP. The subsequent undocking of AMP or tRNA then proceeds along thermodynamically competitive pathways. Release of the tRNA acceptor stem is further accelerated by the deprotonation of the alpha-ammonium group on the charging amino acid. The proposed general base is Glu41
-
purified recombinant enzyme in complex with tRNAGlu, hanging-drop vapour diffusion method, precipitant solution contains 37 mM Na-MOPS, pH 6.7, 22% PEG 1500, 37 mM ammonium sulfate, 1% 2-methyl-2,4-pentanediol, 10 mM MgCl2, 5 mM 2-mercaptoethanol, X-ray diffraction structure determination at 2.4 A resolution, and analysis
P27000
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
42
-
-
stable up to, without addition of substrate
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 1C per min, without addition of substrate
65
-
-
8 h, no loss of activity, in absence of substrate
65
-
-
9 h, 30% loss of activity
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
additional information
-
-
46000 MW polypeptide stabilizes the enzyme against heat inactivation
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
tRNAGlu in presence of Mg2+ protects against heat inactivation, Mg2+ alone is much less effective
-
Mycobacterium tuberculosis GluRS is significantly more sensitive than the Escherichia coli form to tryptic and chymotryptic limited proteolysis. Chymotrypsin-sensitive sites are found in the predicted tRNA stem contact domain next to the ATP binding site. Enzyme is fully protected from proteolysis by ATP and glutamol-AMP
-, P0A636
no stabilization by various sulfydryl reagents, glycerol and glutamic acid
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, 10 mM Tris-HCl, pH 8.0, 1 mM MgCl2, 20 mM 2-mercaptoethanol, 0.1 mM PMSF, 50% glycerol, stable for several months
-
-20C or -70C, 20 mM sodium Hepes, pH 7.2, 0.1 mM EDTA, 0.5 mM DTT, 65% glycerol, stable for at least 1 year
-
85% loss of activity after 24 h
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant GST-GluRS1 by glutathione affinity chromatography, followed by removal of GST
-
50fold, recombinant enzyme from overexpression in Bacillus subtilis, to homogeneity
-
-
Caesalpinia bondue
-
partial, recombinant enzyme from Escherichia coli with or without N-terminal His-tag
-
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
-
six-h procedure
-
recombinant enzyme
-, P0A636
recombinant His-tagged GluRS from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
-
recombinant His6-tagged GluRS-N from Escherichia coli strain BL21(DE3) to homogeneity by nickel affinity chromatography and gel filtration
-
recombinant TM1351 from Escherichia coli strain Rosetta2 (DE3) by anion exchange chromatography and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli; gene gltX1 and gltX2, expression in Escherichia coli temperature-sensitive mutant strain
-
overexpression of GST-GluRS1
-
overexpression of GST-tagged isozyme GluRS1 in Escherichia coli strain Bl21(DE3), co-expression with Glu-tRNA reductase
-
cloning and overexpression in Escherichia coli, coexpression of recombinant tRNA mutant variants
-
DNA and amino acid sequence determination and analysis, full length gene, single gene, expression in Escherichia coli BL21 as S-tagged enzyme
O82462
cloning and overexpression in Escherichia coli, coexpression of recombinant tRNA mutant variants
-
gene gltX is encoded in an operon comprising the genes gltX, cysE, and cysS, after transcription the polycistronic mRNA is cleaved into the gltX transcript and the cysE-cysS transcript due to an internal signal structure and probably by self-cleavage, analysis, the gltX mRNA may be stabilized by secondary structures at the 3'-end and the 5'-end, expression in Escherichia coli and in vitro transcription
-
gene gltX mutant, mutant enzyme expression in Escherichia coli temperature-sensitive mutant strain
-
gene gltX, expression in Escherichia coli is lethal, but a modulated enzyme, lacking a part of the tRNA acceptor stem binding domain, can be recombinantly expressed in Escherichia coli DH5alpha, overexpression in Bacillus subtilis
-
overexpression of the enzyme in Escherichia coli with or without N-terminal His-tag
-
expression of His-tagged ERS in strain BL21(DE3)
-
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)
-
gene gltX2, expression in Escherichia coli temperature-sensitive mutant strain
-
expression of C-terminally His-tagged, full-length EPRS
-
cloning and overexpression in Escherichia coli, coexpression of recombinant tRNA mutant variants
Methanocaldococcus jannaschii, Methanococcus thermoautotrophicum, Methanosarcina mazei
-
expression in Escherichia coli
-, P0A636
cloning and overexpression in Escherichia coli, coexpression of recombinant tRNA mutant variants
-
expression of His6-tagged GluRS-N, comprising residues 1-197, 17-207 and 1-207, in Escherichia coli strain BL21(DE3)
-
expression of wild-type and mutant His-tagged GluRSs in Escherichia coli strain BL21(DE3)
-
gene gltX, DNA and amino acid sequence determination and analysis, expression in Escherichia coli
-
cloning and overexpression in Escherichia coli, coexpression of recombinant tRNA mutant variants
-
TM1351, sequence comparisons, expression in Escherichia coli strain Rosetta2(DE3)
-
overexpression in Escherichia coli
-
phylogenetic and expression analysis, the tnpA gene of the IS element and gene gltX are co-transcribed and their expression is transiently upregulated upon retrieval of the ammonium source irrespective of whether nitrate or no nitrogen source are available, structural elements of the promoter directing the expression of the tnpA-gltX operon are localized within the IS
Tolypothrix sp.
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
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
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ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Q373R
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substrate specificity restricted to tRNAGlu compared to the wild-type which also accepts tRNAGln
R358Q
P27000
site-directed mutagenesis, exchange of the Arg residue results in a mutant that no longer discriminates between tRNAGlu and tRNAGln anticodons YUC and YUG, respectively
additional information
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design of orthogonal tRNAs with sequences derived from archaeal tRNAs, 3 forms: 1. AQ(GU), i.e. archaeal glutaminyl with GU base pair at position 10-26, 2. AE(GU), i.e. archaeal glutamyl with GU base pair at position 10-28, 3. AE(GC), i.e. archaeal glutamyl with GC base pair at position 10-28
additional information
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construction and expression in Escherichia coli of mutant forms of Escherichia coli tRNAGln2 and tRNAGln1 and of Bacillus subtilis tRNAGln
H129Q
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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
additional information
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construction and overexpression of tRNAGlu variants, the enzyme shows altered activity with the tRNA mutants, structural characterization
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. 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
additional information
P04805
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
Methanocaldococcus jannaschii, Methanococcus thermoautotrophicum, Methanosarcina mazei
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design of orthogonal tRNAs with sequences derived from archaeal tRNAs, 3 forms: 1. AQ(GU), i.e. archaeal glutaminyl with GU base pair at position 10-26, 2. AE(GU), i.e. archaeal glutamyl with GU base pair at position 10-28, 3. AE(GC), i.e. archaeal glutamyl with GC base pair at position 10-28
additional information
-, P0A636
protein is not toxic when overproduced in Escherichia coli cells indicating that it does not catalyze the mischarging of Escherichia coli tRNAGln with l-Glu and that GluRS /tRNAGln recognition is species specific
additional information
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design of orthogonal tRNAs with sequences derived from archaeal tRNAs, 3 forms: 1. AQ(GU), i.e. archaeal glutaminyl with GU base pair at position 10-26, 2. AE(GU), i.e. archaeal glutamyl with GU base pair at position 10-28, 3. AE(GC), i.e. archaeal glutamyl with GC base pair at position 10-28
additional information
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construction of truncated enzyme GluRS-N, comprising residues 1-197, 17-207 and 1-207
additional information
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design of orthogonal tRNAs with sequences derived from archaeal tRNAs, 3 forms: 1. AQ(GU), i.e. archaeal glutaminyl with GU base pair at position 10-26, 2. AE(GU), i.e. archaeal glutamyl with GU base pair at position 10-28, 3. AE(GC), i.e. archaeal glutamyl with GC base pair at position 10-28
additional information
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mutant enzymes with higher Km and lower turnover numbers
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
drug development
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design of better inhibitors specific for bacterial GluRSs, which are promising targets for antimicrobial therapy