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Literature summary extracted from

  • Zhou, X.L.; Zhu, B.; Wang, E.D.
    The CP2 domain of leucyl-tRNA synthetase is crucial for amino acid activation and post-transfer editing (2008), J. Biol. Chem., 283, 36608-36616.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.1.1.4 expressed in Escherichia coli as a His-tagged fusion protein Giardia intestinalis

Protein Variants

EC Number Protein Variants Comment Organism
6.1.1.4 D588A kcat/Km: 0.57 (ATP), 0.37 (Leu), mutant displays lower amino acid activation and aminoacylation activities than wild-type Giardia intestinalis
6.1.1.4 D603A kcat/Km: 0.91 (ATP), 0.94 (Leu), similar activity compared to wild-type Giardia intestinalis
6.1.1.4 DELTA581-617 deletion of the CP2 domain, an insertion domain called connective peptide 2, of Giardia lamblia LeuRS shows that the CP2 domain is indispensable for amino acid activation, post-transfer editing and contributes to LeuRS-tRNALeu binding affinity. CP2 domain of Pyrococcus horikoshii LeuRS but not that of Escherichia coli LeuRS can partially restore amino acid activation and post-transfer editing functions suggesting that the functions of the CP2 domain are dependent on its location in the primary sequence of LeuRS Giardia intestinalis
6.1.1.4 K587A kcat/Km: 0.43 (ATP), 0.42 (Leu), mutant displays lower amino acid activation and aminoacylation activities than wild-type Giardia intestinalis
6.1.1.4 K606A mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities Giardia intestinalis
6.1.1.4 K606D kcat/Km: 0.86 (ATP), 0.83 (Leu), similar leucine activation and post-transfer editing activities compared to wild-type Giardia intestinalis
6.1.1.4 K606E kcat/Km: 0.90 (ATP), 0.99 (Leu), no difference in leucine activation and post-transfer editing activities compared to wild-type Giardia intestinalis
6.1.1.4 K606L kcat/Km: 0.85 (ATP), 0.98 (Leu), similar leucine activation and post-transfer editing activities compared to wild-type Giardia intestinalis
6.1.1.4 K606R kcat/Km: 0.98 (ATP), 0.99 (Leu), no difference in leucine activation and post-transfer editing activities compared to wild-type Giardia intestinalis
6.1.1.4 W586A mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities Giardia intestinalis
6.1.1.4 Y581A mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities Giardia intestinalis
6.1.1.4 Y581E mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity Giardia intestinalis
6.1.1.4 Y581F mutant shows some but significantly reduced leucine activation activity its post-transfer editing activity is similar to wild-type Giardia intestinalis
6.1.1.4 Y581K mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity Giardia intestinalis
6.1.1.4 Y581S mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity Giardia intestinalis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.1.1.4 additional information
-
 additional information kcat/Km: 1 (ATP, wild-type), 1 (Leu, wild-type), 0.43 (ATP, mutant K587A), 0.42 (Leu, mutant K587A), 0.57 (ATP, mutant K588A), 0.37 (Leu, mutant K588A), 0.91 (ATP, mutant D603A), 0.94 (Leu, mutant D603A), 0.98 (ATP, mutant K606R), 0.99 (Leu, mutant K606R), 0.9 (ATP, mutant K606E), 0.99 (Leu, mutant K606E), 0.85 (ATP, mutant K606L), 0.98 (Leu, mutant K606L), 0.86 (ATP, mutant K606D), 0.83 (Leu, mutant K606D) Giardia intestinalis

Organism

EC Number Organism UniProt Comment Textmining
6.1.1.4 Giardia intestinalis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.1.1.4 using Ni-NTA chromatography Giardia intestinalis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.1.1.4 ATP + L-isoleucine + tRNALeu
-
 Giardia intestinalis AMP + diphosphate + L-isoleucyl-tRNALeu
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.1.1.4 GlLeuRS
-
 Giardia intestinalis
6.1.1.4 Leucyl-tRNA synthetase
-
 Giardia intestinalis

Temperature Optimum [°C]

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

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.1.1.4 8.2
-
 assay at Giardia intestinalis