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Literature summary extracted from

  • Yan, W.; Ye, Q.; Tan, M.; Chen, X.; Eriani, G.; Wang, E.D.
    Modulation of aminoacylation and editing properties of leucyl-tRNA synthetase by a conserved structural module (2015), J. Biol. Chem., 290, 12256-12267 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.1.1.4 recombinant expression of wild-type and mutant enzymes Homo sapiens
6.1.1.4 recombinant expression of wild-type and mutant enzymes Escherichia coli
6.1.1.4 recombinant expression of wild-type and mutant enzymes Mesomycoplasma mobile
6.1.1.4 recombinant expression of wild-type and mutant enzymes Aquifex aeolicus
6.1.1.4 recombinant expression of wild-type and mutant enzymes Pyrococcus horikoshii

Protein Variants

EC Number Protein Variants Comment Organism
6.1.1.4 K452A site-directed mutagenesis, the mutation has only a minimal effect on aminoacylation activity, the Km values is not significantly altered compared to wild-type Mesomycoplasma mobile
6.1.1.4 K452E site-directed mutagenesis, the mutation has only a minimal effect on aminoacylation activity, the Km values is not significantly altered compared to wild-type Mesomycoplasma mobile
6.1.1.4 K671A site-directed mutagenesis, the mutation does no affect the catalytic efificiency Escherichia coli
6.1.1.4 K692A site-directed mutagenesis, the mutation has no effect on tRNA charging activity Pyrococcus horikoshii
6.1.1.4 K696A site-directed mutagenesis, the mutant shows a highly reduced kcat value compared to wild-type, while the Km value is 3fold increased Pyrococcus horikoshii
6.1.1.4 K699A site-directed mutagenesis, the mutation has no effect on tRNA charging activity Pyrococcus horikoshii
6.1.1.4 additional information a helix alpha3-deletion mutant is inactive Escherichia coli
6.1.1.4 R456A site-directed mutagenesis, 75% reduced catalytic efficiency compared to wild-type, the Km values is not significantly altered Mesomycoplasma mobile
6.1.1.4 R456E site-directed mutagenesis, 79% reduced catalytic efficiency compared to wild-type, the Km values is not significantly altered Mesomycoplasma mobile
6.1.1.4 R668A site-directed mutagenesis, the mutant shows 77% reduced catalytic efficiency compared to wild-type, the rate of AMP formation is decreased compared to the wild-type Escherichia coli
6.1.1.4 R668A/R672A site-directed mutagenesis, the mutant shows 93.6% reduced catalytic efficiency compared to wild-type, the rate of AMP formation is decreased compared to the wild-type Escherichia coli
6.1.1.4 R668E site-directed mutagenesis, the mutant shows 95% reduced catalytic efficiency compared to wild-type, the rate of AMP formation is decreased compared to the wild-type Escherichia coli
6.1.1.4 R668E/R672E site-directed mutagenesis, the mutant shows 98.6% reduced catalytic efficiency compared to wild-type. But the almost inactive mutant exhibits intact Leu activation activity comparable with the wild-type enzyme Escherichia coli
6.1.1.4 R672A site-directed mutagenesis, the rate of AMP formation is decreased compared to the wild-type Escherichia coli
6.1.1.4 R672E site-directed mutagenesis, the rate of AMP formation is decreased compared to the wild-type Escherichia coli
6.1.1.4 R698A site-directed mutagenesis, the mutation has no effect on tRNA charging activity Pyrococcus horikoshii
6.1.1.4 R703A site-directed mutagenesis, kcat of mutant PhLeuRSR703A is much lower than that of wild-type PhLeuRS Pyrococcus horikoshii
6.1.1.4 R766A site-directed mutagenesis, the mutation decreases the kcat/Km value to less than 10% that of the wild-type enzyme hcLeuRS Homo sapiens
6.1.1.4 R94A site-directed mutagenesis, mutating Arg94 to Ala decreases kcat/Km values to 34% of that of wild-type AaLeuRS Aquifex aeolicus
6.1.1.4 R94A/R98A site-directed mutagenesis, 83% reduced catalytic efficiency compared to wild-type Aquifex aeolicus
6.1.1.4 R94E site-directed mutagenesis, mutating Arg94 to Glu decreases kcat/Km values to 22% of that of wild-type AaLeuRS Aquifex aeolicus
6.1.1.4 R94E/R98E site-directed mutagenesis, the rate of AMP formation is decreased compared to the wild-type Aquifex aeolicus
6.1.1.4 R98A site-directed mutagenesis, the mutation does not alter the catalytic efficiency Aquifex aeolicus
6.1.1.4 R98E site-directed mutagenesis, the rate of AMP formation is decreased compared to the wild-type Aquifex aeolicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.1.1.4 additional information
-
 additional information dissociation constants of LeuRS and mutants from Escherichia coli for their cognate tRNAs. Reaction kinetics of EcLeuRS for Mycoplasma mobile MmtRNAUAALeu and mutant variants, kinetic constants of EcLeuRS, chimeric LeuRS and their mutants for tRNALeu in aminoacylation reaction and for AMP formation in the presence of Nva and MmtRNACAA, detailed overview Escherichia coli
6.1.1.4 additional information
-
 additional information dissociation constants of LeuRS and mutants from human cytoplasm for their cognate tRNAs Homo sapiens
6.1.1.4 0.00012
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant R703A Pyrococcus horikoshii
6.1.1.4 0.0003
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant wild-type enzyme Aquifex aeolicus
6.1.1.4 0.00031
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R98A Aquifex aeolicus
6.1.1.4 0.00039
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R98E Aquifex aeolicus
6.1.1.4 0.00044
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K692A Pyrococcus horikoshii
6.1.1.4 0.00052
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K698A Pyrococcus horikoshii
6.1.1.4 0.0006
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K699A Pyrococcus horikoshii
6.1.1.4 0.00065
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K696A Pyrococcus horikoshii
6.1.1.4 0.00074
-
 tRNACAGLeu pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens
6.1.1.4 0.00075
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94A Aquifex aeolicus
6.1.1.4 0.00081
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94A/R98A Aquifex aeolicus
6.1.1.4 0.00081
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94E Aquifex aeolicus
6.1.1.4 0.00088
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94E/R98E Aquifex aeolicus
6.1.1.4 0.0015
-
 tRNAUAALeu pH 7.8, 30°C, recombinant wild-type enzyme Mesomycoplasma mobile
6.1.1.4 0.0016
-
 tRNAUAALeu pH 7.5, 37°C, recombinant wild-type enzyme Escherichia coli
6.1.1.4 0.0017
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K452A Mesomycoplasma mobile
6.1.1.4 0.0017
-
 tRNACAGLeu pH 7.8, 37°C, recombinant mutant R668A Homo sapiens
6.1.1.4 0.0019
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K452E Mesomycoplasma mobile
6.1.1.4 0.002
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant wild-type enzyme Pyrococcus horikoshii
6.1.1.4 0.0022
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant wild-type enzyme Escherichia coli
6.1.1.4 0.0022
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668A/R672A Escherichia coli
6.1.1.4 0.0023
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K456A Mesomycoplasma mobile
6.1.1.4 0.0024
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant K671A Escherichia coli
6.1.1.4 0.0024
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R672A Escherichia coli
6.1.1.4 0.0025
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K456E Mesomycoplasma mobile
6.1.1.4 0.0028
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668A Escherichia coli
6.1.1.4 0.0028
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R672E Escherichia coli
6.1.1.4 0.0054
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668E Escherichia coli
6.1.1.4 0.0058
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668E/R672E Escherichia coli

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
6.1.1.4 cytoplasm
-
 Homo sapiens 5737
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.1.1.4 Mg2+ required Homo sapiens
6.1.1.4 Mg2+ required Escherichia coli
6.1.1.4 Mg2+ required Mesomycoplasma mobile
6.1.1.4 Mg2+ required Aquifex aeolicus
6.1.1.4 Mg2+ required Pyrococcus horikoshii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.1.1.4 ATP + L-leucine + tRNALeu Homo sapiens
-
 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu Escherichia coli
-
 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu Mesomycoplasma mobile
-
 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu Aquifex aeolicus
-
 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu Pyrococcus horikoshii
-
 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu Mesomycoplasma mobile ATCC 43663 / 163K / NCTC 11711
-
 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu Pyrococcus horikoshii ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3
-
 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.1.1.4 Aquifex aeolicus O66680 AND O67646 alpha- and beta-subunit
-
6.1.1.4 Escherichia coli P07813
-
-
6.1.1.4 Homo sapiens Q9P2J5
-
-
6.1.1.4 Mesomycoplasma mobile Q6KHA5
-
-
6.1.1.4 Mesomycoplasma mobile ATCC 43663 / 163K / NCTC 11711 Q6KHA5
-
-
6.1.1.4 Pyrococcus horikoshii O58698
-
-
6.1.1.4 Pyrococcus horikoshii ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3 O58698
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.1.1.4 recombinant wild-type and mutant enzymes Homo sapiens
6.1.1.4 recombinant wild-type and mutant enzymes Escherichia coli
6.1.1.4 recombinant wild-type and mutant enzymes Mesomycoplasma mobile
6.1.1.4 recombinant wild-type and mutant enzymes Aquifex aeolicus
6.1.1.4 recombinant wild-type and mutant enzymes Pyrococcus horikoshii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.1.1.4 ATP + L-leucine + tRNACAALeu Mycoplasma mobile tRNACAALeu (MmtRNACAALeu) and mutat derivatives Mesomycoplasma mobile AMP + diphosphate + L-leucyl-tRNAUAALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNACAALeu Mycoplasma mobile tRNACAALeu (MmtRNACAALeu) and mutat derivatives Mesomycoplasma mobile ATCC 43663 / 163K / NCTC 11711 AMP + diphosphate + L-leucyl-tRNAUAALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNACAGLeu human cytoplasmic tRNACAGLeu (hctRNACAG) Homo sapiens AMP + diphosphate + L-leucyl-tRNACAGLeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNAGAGLeu Aquifex aeolicus tRNAGAGLeu (AatRNAGAGLeu) Aquifex aeolicus AMP + diphosphate + L-leucyl-tRNAGAGLeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNAGAGLeu Escherichia coli tRNAGAGLeu (Ect-RNAGAGLeu) Escherichia coli AMP + diphosphate + L-leucyl-tRNAGAGLeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNAGAGLeu Pyrococcus horikoshii tRNAGAGLeu (PhtRNAGAGLeu) Pyrococcus horikoshii AMP + diphosphate + L-leucyl-tRNAGAGLeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNAGAGLeu Pyrococcus horikoshii tRNAGAGLeu (PhtRNAGAGLeu) Pyrococcus horikoshii ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3 AMP + diphosphate + L-leucyl-tRNAGAGLeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu
-
 Homo sapiens AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu
-
 Escherichia coli AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu
-
 Mesomycoplasma mobile AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu
-
 Aquifex aeolicus AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu
-
 Pyrococcus horikoshii AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu
-
 Mesomycoplasma mobile ATCC 43663 / 163K / NCTC 11711 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNALeu
-
 Pyrococcus horikoshii ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNAUAALeu Mycoplasma mobile tRNAUAALeu (MmtRNAUAALeu) and mutat derivatives Mesomycoplasma mobile AMP + diphosphate + L-leucyl-tRNACAALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNAUAALeu Mycoplasma mobile tRNAUAALeu (MmtRNAUAALeu) and mutat derivatives Mesomycoplasma mobile ATCC 43663 / 163K / NCTC 11711 AMP + diphosphate + L-leucyl-tRNACAALeu
-
 ?
6.1.1.4 ATP + L-leucine + tRNAUAALeu Mycoplasma mobile MmtRNAUAALeu (Mmt-RNAUAALeu) Escherichia coli AMP + diphosphate + L-leucyl-tRNAUAALeu
-
 ?

Subunits

EC Number Subunits Comment Organism
6.1.1.4 heterodimer
-
 Aquifex aeolicus

Synonyms

EC Number Synonyms Comment Organism
6.1.1.4 AaLeuRS
-
 Aquifex aeolicus
6.1.1.4 EcLeuRS
-
 Escherichia coli
6.1.1.4 HcleuRS
-
 Homo sapiens
6.1.1.4 Leucyl-tRNA synthetase
-
 Homo sapiens
6.1.1.4 Leucyl-tRNA synthetase
-
 Escherichia coli
6.1.1.4 Leucyl-tRNA synthetase
-
 Mesomycoplasma mobile
6.1.1.4 Leucyl-tRNA synthetase
-
 Aquifex aeolicus
6.1.1.4 Leucyl-tRNA synthetase
-
 Pyrococcus horikoshii
6.1.1.4 LeuRS
-
 Homo sapiens
6.1.1.4 LeuRS
-
 Escherichia coli
6.1.1.4 LeuRS
-
 Mesomycoplasma mobile
6.1.1.4 LeuRS
-
 Aquifex aeolicus
6.1.1.4 LeuRS
-
 Pyrococcus horikoshii
6.1.1.4 leuS
-
 Mesomycoplasma mobile
6.1.1.4 leuS
-
 Pyrococcus horikoshii
6.1.1.4 MmLeuRS
-
 Mesomycoplasma mobile
6.1.1.4 PhLeuRS
-
 Pyrococcus horikoshii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.1.1.4 30
-
 assay at Mesomycoplasma mobile
6.1.1.4 37
-
 assay at Homo sapiens
6.1.1.4 37
-
 assay at Escherichia coli
6.1.1.4 37
-
 assay at Pyrococcus horikoshii
6.1.1.4 65
-
 assay at Aquifex aeolicus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.1.1.4 0.0055
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant R703A Pyrococcus horikoshii
6.1.1.4 0.017
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K696A Pyrococcus horikoshii
6.1.1.4 0.02
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant wild-type enzyme Pyrococcus horikoshii
6.1.1.4 0.041
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K692A Pyrococcus horikoshii
6.1.1.4 0.05
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K699A Pyrococcus horikoshii
6.1.1.4 0.067
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K698A Pyrococcus horikoshii
6.1.1.4 0.18
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668E/R672E Escherichia coli
6.1.1.4 0.19
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94E/R98E Aquifex aeolicus
6.1.1.4 0.31
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668A/R672A Escherichia coli
6.1.1.4 0.53
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R672E Escherichia coli
6.1.1.4 0.56
-
 tRNACAGLeu pH 7.8, 37°C, recombinant mutant R668A Homo sapiens
6.1.1.4 0.59
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668E Escherichia coli
6.1.1.4 0.67
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94A/R98A Aquifex aeolicus
6.1.1.4 0.85
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K456E Mesomycoplasma mobile
6.1.1.4 0.89
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94E Aquifex aeolicus
6.1.1.4 1
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R98E Aquifex aeolicus
6.1.1.4 1.3
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K456A Mesomycoplasma mobile
6.1.1.4 1.3
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94A Aquifex aeolicus
6.1.1.4 1.4
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668A Escherichia coli
6.1.1.4 1.5
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R98A Aquifex aeolicus
6.1.1.4 1.5
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant wild-type enzyme Aquifex aeolicus
6.1.1.4 2.6
-
 tRNACAGLeu pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens
6.1.1.4 2.8
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K452E Mesomycoplasma mobile
6.1.1.4 3.2
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K452A Mesomycoplasma mobile
6.1.1.4 3.3
-
 tRNAUAALeu pH 7.8, 30°C, recombinant wild-type enzyme Mesomycoplasma mobile
6.1.1.4 4.2
-
 tRNAUAALeu pH 7.5, 37°C, recombinant wild-type enzyme Escherichia coli
6.1.1.4 4.8
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant K671A Escherichia coli
6.1.1.4 4.8
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R672A Escherichia coli
6.1.1.4 4.9
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant wild-type enzyme Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.1.1.4 7.5
-
 assay at Escherichia coli
6.1.1.4 7.8
-
 assay at Homo sapiens
6.1.1.4 7.8
-
 assay at Mesomycoplasma mobile
6.1.1.4 7.8
-
 assay at Aquifex aeolicus
6.1.1.4 7.8
-
 assay at Pyrococcus horikoshii

Cofactor

EC Number Cofactor Comment Organism Structure
6.1.1.4 ATP
-
 Homo sapiens
6.1.1.4 ATP
-
 Escherichia coli
6.1.1.4 ATP
-
 Mesomycoplasma mobile
6.1.1.4 ATP
-
 Aquifex aeolicus
6.1.1.4 ATP
-
 Pyrococcus horikoshii

General Information

EC Number General Information Comment Organism
6.1.1.4 evolution based on sequence homology and the structures of the catalytic active sites, aaRSs are divided into two classes of 10 members each. Class I synthetases are further divided into three subclasses, a, b, and c, according to sequence homology. Leucyl-tRNA synthetase (LeuRS) belongs to class I aaRSs that include a typical Rossmann dinucleotide-binding fold active site architecture with the signature sequence modules HIGH and KMSKS. According to evolutionary models, the primitive catalytic core is extended by the insertion and/or fusion of additional domains (also called modules) in LeuRSs, most of which have inserted a large connective polypeptide 1 (CP1) domain that is responsible for amino acid editing. To ensure translation accuracy, LeuRSs have evolved a mechanism to remove aminoacyl AMP (aa-AMP, pre-transfer editing) and aa-tRNA (post-transfer editing). Sequence comparisons of the stem contact-fold domain (SC-fold) involved in editing, basic residues on helix alpha3 of the SC-fold are critical for catalytic efficiency, overview Homo sapiens
6.1.1.4 evolution based on sequence homology and the structures of the catalytic active sites, aaRSs are divided into two classes of 10 members each. Class I synthetases are further divided into three subclasses, a, b, and c, according to sequence homology. Leucyl-tRNA synthetase (LeuRS) belongs to class I aaRSs that include a typical Rossmann dinucleotide-binding fold active site architecture with the signature sequence modules HIGH and KMSKS. According to evolutionary models, the primitive catalytic core is extended by the insertion and/or fusion of additional domains (also called modules) in LeuRSs, most of which have inserted a large connective polypeptide 1 (CP1) domain that is responsible for amino acid editing. To ensure translation accuracy, LeuRSs have evolved a mechanism to remove aminoacyl AMP (aa-AMP, pre-transfer editing) and aa-tRNA (post-transfer editing). Sequence comparisons of the stem contact-fold domain (SC-fold) involved in editing, basic residues on helix alpha3 of the SC-fold are critical for catalytic efficiency, overview Aquifex aeolicus
6.1.1.4 evolution based on sequence homology and the structures of the catalytic active sites, aaRSs are divided into two classes of 10 members each. Class I synthetases are further divided into three subclasses, a, b, and c, according to sequence homology. Leucyl-tRNA synthetase (LeuRS) belongs to class I aaRSs that include a typical Rossmann dinucleotide-bindingfold active site architecture with the signature sequence modules HIGH and KMSKS. According to evolutionary models, the primitive catalytic core is extended by the insertion and/or fusion of additional domains (also called modules) in LeuRSs, most of which have inserted a large connective polypeptide 1 (CP1) domain that is responsible for amino acid editing. To ensure translation accuracy, LeuRSs have evolved a mechanism to remove aminoacyl AMP (aa-AMP, pre-transfer editing) and aa-tRNA (post-transfer editing). Although post-transfer editing is carried out by the CP1 domain in most LeuRSs, this domain has been naturally deleted in LeuRS from Mycoplasma mobile (MmLeuRS). Sequence comparisons of the stem contact-fold domain (SC-fold) involved in editing, basic residues on helix alpha3 of the SC-fold are critical for catalytic efficiency, overview Mesomycoplasma mobile
6.1.1.4 evolution based on sequence homology and the structures of the catalytic active sites, aaRSs are divided into two classes of 10 members each. Class I synthetases are further divided into three subclasses, a, b, and c, according to sequence homology. Leucyl-tRNA synthetase (LeuRS) belongs to class I aaRSs that include a typical Rossmann dinucleotide-bindingfold active site architecture with the signature sequence modules HIGH and KMSKS. According to evolutionary models, the primitive catalytic core is extended by the insertion and/or fusion of additional domains (also called modules) in LeuRSs, most of which have inserted a large connective polypeptide 1 (CP1) domain that is responsible for amino acid editing. To ensure translation accuracy, LeuRSs have evolved a mechanism to remove aminoacyl AMP (aa-AMP, pre-transfer editing) and aa-tRNA (post-transfer editing). Sequence comparison of the EcLeuRS stem contact-fold domain (SC-fold) with editing-deficient enzymes suggests that key residues of this module have evolved an adaptive strategy to follow the editing functions of LeuRS, basic residues on helix alpha3 of the SC-fold are critical for catalytic efficiency, overview Escherichia coli
6.1.1.4 evolution based on sequence homology and the structures of the catalytic active sites, aaRSs are divided into two classes of 10 members each. Class I synthetases are further divided into three subclasses, a, b, and c, according to sequence homology. Leucyl-tRNA synthetase (LeuRS) belongs to class I aaRSs that include a typical Rossmann dinucleotide-bindingfold active site architecture with the signature sequence modules HIGH and KMSKS. According to evolutionary models, the primitive catalytic core is extended by the insertion and/or fusion of additional domains (also called modules) in LeuRSs, most of which have inserted a large connective polypeptide 1 (CP1) domain that is responsible for amino acid editing. To ensure translation accuracy, LeuRSs have evolved a mechanism to remove aminoacyl AMP (aa-AMP, pre-transfer editing) and aa-tRNA (post-transfer editing). Sequence comparisons of the stem contact-fold domain (SC-fold) involved in editing, basic residues on helix alpha3 of the SC-fold are critical for catalytic efficiency, overview Pyrococcus horikoshii
6.1.1.4 malfunction mutation of highly conserved basic residues on the third alpha-helix of the KMSKS catalytic loop domain impairs the affinity of LeuRS for the anticodon stem of tRNALeu, which decreases both aminoacylation and editing activities Escherichia coli
6.1.1.4 additional information the KMSKS catalytic loop exhibits alpha-alpha-beta-alpha topology in class Ia and Ib aminoacyl-tRNA synthetases, two glycine residues on the third alpha-helix contribute to flexibility, leucine activation, and editing of LeuRS from Escherichia coli (EcLeuRS), acidic residues on the beta-strand enhance the editing activity of EcLeuRS and sense the size of the tRNALeu D-loop. Incorporation of acidic residues on the beta-strand stimulates the tRNA-dependent editing activity of the chimeric minimalist enzyme Mycoplasma mobile LeuRS fused to the connective polypeptide 1 editing domain and leucine-specific domain from EcLeuRS. Sequence comparison of the EcLeuRS stem contact-fold domain with editing-deficient enzymes suggests that key residues of this module have evolved an adaptive strategy to follow the editing functions of LeuRS. Amino acid residues Arg668 or Arg672 are not involved in the amino acid activation step but rather the second tRNA transfer step Escherichia coli
6.1.1.4 additional information the KMSKS catalytic loop exhibits alpha-alpha-beta-alpha topology in class Ia and Ib aminoacyl-tRNA synthetases. Incorporation of acidic residues on the beta-strand stimulates the tRNA-dependent editing activity of the chimeric minimalist enzyme Mycoplasma mobile LeuRS fused to the connective polypeptide 1 editing domain and leucine-specific domain from EcLeuRS, acidic residues on the beta-strand enhance the editing activity of EcLeuRS and sense the size of the tRNALeu D-loop Mesomycoplasma mobile
6.1.1.4 physiological function aminoacyl-tRNA synthetases (aaRSs) are a large and diverse family of enzymes that catalyze the attachment of amino acids to their cognate tRNAs in a two-step aminoacylation reaction as follows: 1. amino acid activation by ATP hydrolysis to form an aminoacyl-adenylate intermediate, and 2. transfer of the aminoacyl moiety from the intermediate to the cognate tRNA isoacceptor to form aminoacyl-tRNA (aa-tRNA) Homo sapiens
6.1.1.4 physiological function aminoacyl-tRNA synthetases (aaRSs) are a large and diverse family of enzymes that catalyze the attachment of amino acids to their cognate tRNAs in a two-step aminoacylation reaction as follows: 1. amino acid activation by ATP hydrolysis to form an aminoacyl-adenylate intermediate, and 2. transfer of the aminoacyl moiety from the intermediate to the cognate tRNA isoacceptor to form aminoacyl-tRNA (aa-tRNA) Escherichia coli
6.1.1.4 physiological function aminoacyl-tRNA synthetases (aaRSs) are a large and diverse family of enzymes that catalyze the attachment of amino acids to their cognate tRNAs in a two-step aminoacylation reaction as follows: 1. amino acid activation by ATP hydrolysis to form an aminoacyl-adenylate intermediate, and 2. transfer of the aminoacyl moiety from the intermediate to the cognate tRNA isoacceptor to form aminoacyl-tRNA (aa-tRNA) Mesomycoplasma mobile
6.1.1.4 physiological function aminoacyl-tRNA synthetases (aaRSs) are a large and diverse family of enzymes that catalyze the attachment of amino acids to their cognate tRNAs in a two-step aminoacylation reaction as follows: 1. amino acid activation by ATP hydrolysis to form an aminoacyl-adenylate intermediate, and 2. transfer of the aminoacyl moiety from the intermediate to the cognate tRNA isoacceptor to form aminoacyl-tRNA (aa-tRNA) Aquifex aeolicus
6.1.1.4 physiological function aminoacyl-tRNA synthetases (aaRSs) are a large and diverse family of enzymes that catalyze the attachment of amino acids to their cognate tRNAs in a two-step aminoacylation reaction as follows: 1. amino acid activation by ATP hydrolysis to form an aminoacyl-adenylate intermediate, and 2. transfer of the aminoacyl moiety from the intermediate to the cognate tRNA isoacceptor to form aminoacyl-tRNA (aa-tRNA) Pyrococcus horikoshii

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.1.1.4 30
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668E/R672E Escherichia coli
6.1.1.4 30
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K696A Pyrococcus horikoshii
6.1.1.4 50
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant R703A Pyrococcus horikoshii
6.1.1.4 80
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K699A Pyrococcus horikoshii
6.1.1.4 90
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K692A Pyrococcus horikoshii
6.1.1.4 110
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant wild-type enzyme Pyrococcus horikoshii
6.1.1.4 110
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668E Escherichia coli
6.1.1.4 130
-
 tRNAGAGLeu pH 7.8, 37°C, recombinant mutant K698A Pyrococcus horikoshii
6.1.1.4 140
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668A/R672A Escherichia coli
6.1.1.4 190
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R672E Escherichia coli
6.1.1.4 220
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94E/R98E Aquifex aeolicus
6.1.1.4 330
-
 tRNACAGLeu pH 7.8, 37°C, recombinant mutant R668A Homo sapiens
6.1.1.4 340
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K456E Mesomycoplasma mobile
6.1.1.4 500
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R668A Escherichia coli
6.1.1.4 600
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K456A Mesomycoplasma mobile
6.1.1.4 830
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94A/R98A Aquifex aeolicus
6.1.1.4 1100
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94E Aquifex aeolicus
6.1.1.4 1500
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K452E Mesomycoplasma mobile
6.1.1.4 1700
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R94A Aquifex aeolicus
6.1.1.4 1900
-
 tRNAUAALeu pH 7.8, 30°C, recombinant mutant K452A Mesomycoplasma mobile
6.1.1.4 2000
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant K671A Escherichia coli
6.1.1.4 2000
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant mutant R672A Escherichia coli
6.1.1.4 2200
-
 tRNAUAALeu pH 7.8, 30°C, recombinant wild-type enzyme Mesomycoplasma mobile
6.1.1.4 2230
-
 tRNAGAGLeu pH 7.5, 37°C, recombinant wild-type enzyme Escherichia coli
6.1.1.4 2600
-
 tRNAUAALeu pH 7.5, 37°C, recombinant wild-type enzyme Escherichia coli
6.1.1.4 2600
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R98E Aquifex aeolicus
6.1.1.4 3500
-
 tRNACAGLeu pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens
6.1.1.4 4800
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant mutant R98A Aquifex aeolicus
6.1.1.4 5000
-
 tRNAGAGLeu pH 7.8, 65°C, recombinant wild-type enzyme Aquifex aeolicus