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Literature summary extracted from

  • Du, X.; Wang, E.D.
    E292 is important for the aminoacylation activity of Escherichia coli leucyl-tRNA synthetase (2003), J. Protein Chem., 22, 71-76.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.1.1.4 overexpression of wild-type and mutants Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
6.1.1.4 E292A unaltered specific activity in amino acid activation reaction, 61% reduced aminoacylation activity compared to the wild-type Escherichia coli
6.1.1.4 E292D unaltered specific activity in amino acid activation reaction, 53% reduced aminoacylation activity compared to the wild-type Escherichia coli
6.1.1.4 E292F unaltered specific activity in amino acid activation reaction, 60% reduced aminoacylation activity compared to the wild-type Escherichia coli
6.1.1.4 E292K unaltered specific activity in amino acid activation reaction, 85% reduced aminoacylation activity compared to the wild-type Escherichia coli
6.1.1.4 E292Q unaltered specific activity in amino acid activation reaction, 54% reduced aminoacylation activity compared to the wild-type Escherichia coli
6.1.1.4 E292S unaltered specific activity in amino acid activation reaction, 66% reduced aminoacylation activity compared to the wild-type Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.1.1.4 ATP at high concentration, the mutants are more sensisitve than the wild-type enzyme Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.1.1.4 0.0012
-
 tRNALeu mutant E292K, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.0021
-
 tRNALeu mutant E292A, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.0024
-
 tRNALeu mutant E292S, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.0025
-
 tRNALeu wild-type enzyme, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.0031
-
 tRNALeu mutant E292Q, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.004
-
 tRNALeu mutants E292D and E292F, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.012
-
 L-leucine mutant E292K, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.013
-
 L-leucine mutant E292S, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.014
-
 L-leucine mutant E292D, E292A, and E292F pH 7.8, 37°C Escherichia coli
6.1.1.4 0.015
-
 L-leucine wild-type enzyme and mutant E292Q, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.08
-
 ATP mutant E292F, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.1
-
 ATP mutants E292Q, E292D, and E292A, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.11
-
 ATP mutants E292D and E292K, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.26
-
 ATP wild-type enzyme, pH 7.8, 37°C Escherichia coli

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 Escherichia coli
-
 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.1.1.4 Escherichia coli
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.1.1.4 recombinant wild-type and mutant enzymes from overproducing strain Escherichia coli

Reaction

EC Number Reaction Comment Organism Reaction ID
6.1.1.4 ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu residue E292 is important for aminoacylation activity Escherichia coli
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.1.1.4 additional information
-
 aminoacid activation activities of wild-type and mutant enzymes Escherichia coli
6.1.1.4 0.05
-
 recombinant mutant E292K, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.12
-
 recombinant mutant E292S, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.14
-
 recombinant mutants E292F and E292A, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.16
-
 recombinant mutant E292Q, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.17
-
 recombinant mutant E292D, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.35
-
 recombinant wild-type enzyme, pH 7.8, 37°C Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.1.1.4 ATP + L-leucine + tRNALeu
-
 Escherichia coli AMP + diphosphate + L-leucyl-tRNALeu
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.1.1.4 Leucine translase
-
 Escherichia coli
6.1.1.4 Leucine--tRNA ligase
-
 Escherichia coli
6.1.1.4 Leucyl-transfer ribonucleate synthetase
-
 Escherichia coli
6.1.1.4 Leucyl-transfer ribonucleic acid synthetase
-
 Escherichia coli
6.1.1.4 Leucyl-transfer RNA synthetase
-
 Escherichia coli
6.1.1.4 Leucyl-tRNA synthetase
-
 Escherichia coli
6.1.1.4 LeuRS
-
 Escherichia coli
6.1.1.4 Synthetase, leucyl-transfer ribonucleate
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.1.1.4 37
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.1.1.4 0.8
-
 ATP mutant E292K, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.8
-
 L-leucine mutant E292K, pH 7.8, 37°C Escherichia coli
6.1.1.4 0.8
-
 tRNALeu mutant E292K, pH 7.8, 37°C Escherichia coli
6.1.1.4 1.5
-
 ATP mutant E292S, pH 7.8, 37°C Escherichia coli
6.1.1.4 1.6
-
 L-leucine mutant E292S, pH 7.8, 37°C Escherichia coli
6.1.1.4 1.6
-
 tRNALeu mutant E292S, pH 7.8, 37°C Escherichia coli
6.1.1.4 1.8
-
 tRNALeu mutant E292A, pH 7.8, 37°C Escherichia coli
6.1.1.4 1.8
-
 ATP mutant E292F, pH 7.8, 37°C Escherichia coli
6.1.1.4 1.8
-
 L-leucine mutant E292F, pH 7.8, 37°C Escherichia coli
6.1.1.4 1.9
-
 L-leucine mutant E292A, pH 7.8, 37°C Escherichia coli
6.1.1.4 1.9
-
 tRNALeu mutant E292Q, pH 7.8, 37°C Escherichia coli
6.1.1.4 2
-
 tRNALeu mutant E292F, pH 7.8, 37°C Escherichia coli
6.1.1.4 2
-
 ATP mutants E292Q and E292A, pH 7.8, 37°C Escherichia coli
6.1.1.4 2.2
-
 ATP mutant E292D, pH 7.8, 37°C Escherichia coli
6.1.1.4 2.2
-
 tRNALeu mutant E292D, pH 7.8, 37°C Escherichia coli
6.1.1.4 2.2
-
 L-leucine mutant E292Q, pH 7.8, 37°C Escherichia coli
6.1.1.4 2.4
-
 L-leucine mutant E292D, pH 7.8, 37°C Escherichia coli
6.1.1.4 4.9
-
 ATP wild-type enzyme, pH 7.8, 37°C Escherichia coli
6.1.1.4 5
-
 tRNALeu wild-type enzyme, pH 7.8, 37°C Escherichia coli
6.1.1.4 5.1
-
 L-leucine wild-type enzyme, pH 7.8, 37°C Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.1.1.4 7.8
-
 assay at Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
6.1.1.4 ATP optimal concentration for the wild-type enzyme is 4 mM, for the mutants 2 mM Escherichia coli