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Literature summary extracted from

  • Charron, C.; Roy, H.; Blaise, M.; Giege, R.; Kern, D.
    Non-discriminating and discriminating aspartyl-tRNA synthetases differ in the anticodon-binding domain (2003), EMBO J., 22, 1632-1643.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.1.1.12 overexpression in Escherichia coli strain JM103 Thermococcus kodakarensis
6.1.1.12 overexpression of wild-type isozyme AspRS2 and seleno-Met isozyme AspRS2 in Escherichia coli Thermus thermophilus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
6.1.1.12 purified wild-type and seleno-Met isozymes AspRS2, vapour phase diffusion from mother liquid: 100 mM CHES buffer, pH 9.5, 200 mM NaCl, 10% w/v PEG 8000, X-ray diffraction structure determination at 2.3 A resolution, structure analysis Thermus thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
6.1.1.12 additional information wild-type discriminating enzyme is engineered to a non-discriminating mutant by site-directed mutagenesis, a L1 loop exchange Thermococcus kodakarensis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.1.1.12 0.00014
-
 tRNAAsp recombinant wild-type enzyme, pH 7.2, 37°C Thermus thermophilus
6.1.1.12 0.0002
-
 tRNAAsp recombinant wild-type enzyme, pH 7.2, 37°C Thermococcus kodakarensis
6.1.1.12 0.0002
-
 tRNAAsn recombinant wild-type enzyme, pH 7.2, 37°C Thermus thermophilus
6.1.1.12 0.0009
-
 tRNAAsn recombinant mutant enzyme, pH 7.2, 37°C Thermococcus kodakarensis
6.1.1.12 0.001
-
 tRNAAsp recombinant mutant enzyme, pH 7.2, 37°C Thermococcus kodakarensis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.1.1.12 ATP + L-aspartate + tRNAAsn Thermococcus kodakarensis 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 AMP + diphosphate + L-aspartyl-tRNAAsn
-
 ?
6.1.1.12 ATP + L-aspartate + tRNAAsn Thermus thermophilus the archaeal AspRS2 enzyme is nondiscriminating, which means that it forms Asp-tRNAAsp and Asp-tRNAAsn AMP + diphosphate + L-aspartyl-tRNAAsn
-
 ?
6.1.1.12 ATP + L-aspartate + tRNAAsp Thermus thermophilus
-
 AMP + diphosphate + L-aspartyl-tRNAAsp
-
 ?
6.1.1.12 ATP + L-aspartate + tRNAAsp Thermococcus kodakarensis
-
 AMP + diphosphate + L-aspartyl-tRNAAsp
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.1.1.12 Thermococcus kodakarensis
-
-
-
6.1.1.12 Thermus thermophilus
-
 isozyme AspRS2
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.1.1.12 recombinant from overexpressing Escherichia coli strain JM103, to homogeneity Thermococcus kodakarensis
6.1.1.12 recombinant wild-type and seleno-Met isozyme 2 from overexpressing Escherichia coli Thermus thermophilus

Reaction

EC Number Reaction Comment Organism Reaction ID
6.1.1.12 ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp determination of structural features for discriminating or nondiscriminating aminoacylation activity Thermus thermophilus
a
6.1.1.12 ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp determination of structural features for discriminating or nondiscriminating aminoacylation activity Thermococcus kodakarensis
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.1.1.12 additional information
-
-
 Thermus thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.1.1.12 ATP + L-aspartate + tRNAAsn
-
 Thermus thermophilus AMP + diphosphate + L-aspartyl-tRNAAsn
-
 ?
6.1.1.12 ATP + L-aspartate + tRNAAsn
-
 Thermococcus kodakarensis AMP + diphosphate + L-aspartyl-tRNAAsn
-
 ?
6.1.1.12 ATP + L-aspartate + tRNAAsn 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 Thermococcus kodakarensis AMP + diphosphate + L-aspartyl-tRNAAsn
-
 ?
6.1.1.12 ATP + L-aspartate + tRNAAsn the archaeal AspRS2 enzyme is nondiscriminating, which means that it forms Asp-tRNAAsp and Asp-tRNAAsn Thermus thermophilus AMP + diphosphate + L-aspartyl-tRNAAsn
-
 ?
6.1.1.12 ATP + L-aspartate + tRNAAsp
-
 Thermus thermophilus AMP + diphosphate + L-aspartyl-tRNAAsp
-
 ?
6.1.1.12 ATP + L-aspartate + tRNAAsp
-
 Thermococcus kodakarensis AMP + diphosphate + L-aspartyl-tRNAAsp
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.1.1.12 Aspartic acid translase
-
 Thermus thermophilus
6.1.1.12 Aspartic acid translase
-
 Thermococcus kodakarensis
6.1.1.12 Aspartyl ribonucleate synthetase
-
 Thermus thermophilus
6.1.1.12 Aspartyl ribonucleate synthetase
-
 Thermococcus kodakarensis
6.1.1.12 aspartyl ribonuleic synthetase
-
 Thermus thermophilus
6.1.1.12 aspartyl ribonuleic synthetase
-
 Thermococcus kodakarensis
6.1.1.12 Aspartyl-transfer ribonucleic acid synthetase
-
 Thermus thermophilus
6.1.1.12 Aspartyl-transfer ribonucleic acid synthetase
-
 Thermococcus kodakarensis
6.1.1.12 Aspartyl-transfer RNA synthetase
-
 Thermus thermophilus
6.1.1.12 Aspartyl-transfer RNA synthetase
-
 Thermococcus kodakarensis
6.1.1.12 Aspartyl-tRNA synthetase
-
 Thermus thermophilus
6.1.1.12 Aspartyl-tRNA synthetase
-
 Thermococcus kodakarensis
6.1.1.12 AspRS
-
 Thermus thermophilus
6.1.1.12 AspRS
-
 Thermococcus kodakarensis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.1.1.12 37
-
 assay at Thermus thermophilus
6.1.1.12 37
-
 assay at Thermococcus kodakarensis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.1.1.12 0.0008
-
 tRNAAsn recombinant mutant enzyme, pH 7.2, 37°C Thermococcus kodakarensis
6.1.1.12 0.0056
-
 tRNAAsn recombinant wild-type enzyme, pH 7.2, 37°C Thermus thermophilus
6.1.1.12 0.0083
-
 tRNAAsp recombinant mutant enzyme, pH 7.2, 37°C Thermococcus kodakarensis
6.1.1.12 0.0117
-
 tRNAAsp recombinant wild-type enzyme, pH 7.2, 37°C Thermococcus kodakarensis
6.1.1.12 0.042
-
 tRNAAsp recombinant wild-type enzyme, pH 7.2, 37°C Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.1.1.12 7.2
-
 assay at Thermus thermophilus
6.1.1.12 7.2
-
 assay at Thermococcus kodakarensis