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Information on EC 6.1.1.12 - aspartate-tRNA ligase and Organism(s) Thermococcus kodakarensis and UniProt Accession Q52428

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Thermococcus kodakarensis
UNIPROT: Q52428 not found.
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Word Map
The taxonomic range for the selected organisms is: Thermococcus kodakarensis
The enzyme appears in selected viruses and cellular organisms
Synonyms
aspartyl-trna synthetase, asprs, dars2, asnrs, mitochondrial aspartyl-trna synthetase, non-discriminating aspartyl-trna synthetase, cytoplasmic aspartyl-trna synthetase, mt-asprs, discriminating asprs, d-asprs, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Aspartic acid translase
-
Aspartyl ribonucleate synthetase
-
aspartyl ribonuleic synthetase
-
Aspartyl-transfer ribonucleic acid synthetase
-
Aspartyl-transfer RNA synthetase
-
Aspartyl-tRNA synthetase
-
discriminating aspartyl-tRNA synthetase
-
Antigen T5
-
-
-
-
Aspartate--tRNA ligase
-
-
-
-
Aspartic acid translase
Aspartyl ribonucleate synthetase
Aspartyl ribonucleic synthetase
-
-
-
-
aspartyl ribonuleic synthetase
-
-
Aspartyl-transfer ribonucleic acid synthetase
Aspartyl-transfer RNA synthetase
Aspartyl-tRNA synthetase
AspRS
discriminating AspRS
-
-
Synthetase, aspartyl-transfer ribonucleate
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
determination of structural features for discriminating or nondiscriminating aminoacylation activity
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
aminoacyl group transfer
-
esterification
-
-
-
-
Aminoacylation
-
-
-
-
aminoacyl group transfer
-
-
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
L-aspartate:tRNAAsp ligase (AMP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY hide
9027-32-1
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
wild-type enzyme shows no activity with tRNAAsn. Mutant enzymes W26H, K85P and W26H/K85P are active with tRNAAsn
-
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
GTP + L-aspartate + tRNAAsp
GMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
UTP + L-aspartate + tRNAAsp
UMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + Asp + tRNAAsn
AMP + diphosphate + aspartyl-tRNAAsn
show the reaction diagram
discriminating AspRS gains the ability to form Asp-tRNAAsn in vitro when the W26H or K85P changes are introduced independently or in combination
-
-
?
ATP + Asp + tRNAAsp
AMP + diphosphate + aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
?
ATP + L-aspartate + tRNAAsn
AMP + diphosphate + L-aspartyl-tRNAAsn
show the reaction diagram
-
the archaeal AspRS2 enzyme is discriminating, which means that it forms only Asp-tRNAAsp and not Asp-tRNAAsn, the L1 loop exchange mutant is rendered non-dicriminating
-
?
ATP + L-aspartate + tRNAAsp
AMP + diphosphate + L-aspartyl-tRNAAsp
show the reaction diagram
-
-
-
?
additional information
?
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
putrescine
enhances activity, optimum concentrations is 0.07 mM
spermidine
enhances activity, optimum concentrations is 0.07 mM
spermine
enhances activity, optimum concentrations is 0.005 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0031 - 0.0091
tRNAAsn
0.0012 - 0.011
tRNAAsp
0.0009 - 0.0091
tRNAAsn
0.0002 - 0.011
tRNAAsp
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.009 - 0.037
tRNAAsn
0.034 - 0.116
tRNAAsp
0.0008 - 0.037
tRNAAsn
0.0083 - 0.116
tRNAAsp
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.9 - 6
tRNAAsn
5.8 - 45
tRNAAsp
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8
pH 5.5: about 60% of maximal activity, pH 8.0: about 65% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
75
optimum temperature shifts from 65°C to 75°C in the presence of polyamine molecules putrescine, spermine or spermidine
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35 - 75
35°C: about 60% of maximal activity, 75°C: about 50% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.23
calculated from sequence
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50000
50893
x * 50893, calculated from sequence
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
2 * 50000, SDS-PAGE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K85P
the wild-type enzyme shows no activity with tRNAAsn, the mutant is active with tRNAAsn
W26H
the wild-type enzyme shows no activity with tRNAAsn, the mutant is active with tRNAAsn
W26H/K85P
the wild-type enzyme shows no activity with tRNAAsn, the mutant is active with tRNAAsn
K85P
discriminating AspRS gains the ability to form Asp-tRNAAsn. Mutation impairs the ability to synthesize Asp-trNAASp in vitr, 8fold increase in KM-value for tRNAAsp
W26H
discriminating AspRS gains the ability to form Asp-tRNAAsn. Mutation causes a 1.5fold decrease in overall catalytic efficiency for Asp-tRNAASp synthesis
W26H/K85P
discriminating AspRS gains the ability to form Asp-tRNAAsn
additional information
-
wild-type discriminating enzyme is engineered to a non-discriminating mutant by site-directed mutagenesis, a L1 loop exchange
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
100
half-life: 3.5 h
85
the thermal melting temperature seems higher than 85°C in the presence of Mg2+. In the absence of Mg 2+, melting temperature decreases to 85°C
90
half-life: 17 h
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme from Escherichia coli
recombinant from overexpressing Escherichia coli strain JM103, to homogeneity
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression as His-tagged protein in Escherichia coli
expression in Escherichia coli
overexpression in Escherichia coli strain JM103
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Fujiwara, S.; Lee, S.; Haruki, M.; Kanaya, S.; Takagi, M.; Imanaka, T.
Unusual enzyme characteristics of aspartyl-tRNA synthetase from hyperthermophilic archaeon Pyrococcus sp. KOD1
FEBS Lett.
394
66-70
1996
Homo sapiens, Pyrococcus sp., Pyrococcus sp. KOD1, Thermococcus kodakarensis (Q52428)
Manually annotated by BRENDA team
Tumbula-Hansen, D.; Feng, L.; Toogood, H.; Stetter, K.O.; Sll, D.
Evolutionary divergence of the archaeal aspartyl-tRNA synthetases into discriminating and nondiscriminating forms
J. Biol. Chem.
277
37184-37190
2002
Methanothermobacter thermautotrophicus, Ferroplasma acidarmanus, Thermococcus kodakarensis (Q52428), Thermococcus kodakarensis
Manually annotated by BRENDA team
Charron, C.; Roy, H.; Blaise, M.; Giege, R.; Kern, D.
Non-discriminating and discriminating aspartyl-tRNA synthetases differ in the anticodon-binding domain
EMBO J.
22
1632-1643
2003
Thermus thermophilus, Thermococcus kodakarensis
Manually annotated by BRENDA team
Feng, L.; Tumbula-Hansen, D.; Toogood, H.; Soll, D.
Expanding tRNA recognition of a tRNA synthetase by a single amino acid change
Proc. Natl. Acad. Sci. USA
100
5676-5681
2003
Thermococcus kodakarensis, Thermococcus kodakarensis (Q52428)
Manually annotated by BRENDA team
Imanaka, T.; Lee, S.; Takagi, M.; Fujiwara, S.
Aspartyl-tRNA synthetase of the hyperthermophilic archaeon Pyrococcus sp. KOD1 has a chimerical structure of eukaryotic and bacterial enzymes
Gene
164
153-156
1995
Thermococcus kodakarensis (Q52428)
Manually annotated by BRENDA team