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Literature summary for 6.1.1.4 extracted from

  • Huang, Q.; Zhou, X.L.; Hu, Q.H.; Lei, H.Y.; Fang, Z.P.; Yao, P.; Wang, E.D.
    A bridge between the aminoacylation and editing domains of leucyl-tRNA synthetase is crucial for its synthetic activity (2014), RNA, 20, 1440-1450 .
    View publication on PubMedView publication on EuropePMC

Protein Variants

Protein Variants Comment Organism
A525S site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
C527E site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
D250A site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
D250E site-directed mutagenesis, the mutant shows slightly altered kinetics and slightly reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
D250N site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
D250R site-directed mutagenesis, inactive mutant Homo sapiens
D252R site-directed mutagenesis, inactive mutant Homo sapiens
D528R site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 85% reduced amino acid activation activity compared to the wild-type enzyme Homo sapiens
G245A site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
G245D site-directed mutagenesis, the mutant shows altered kinetics and 50% reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
G245P site-directed mutagenesis, inactive mutant Homo sapiens
G245R site-directed mutagenesis, the mutant shows altered kinetics and 50% reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
H251D site-directed mutagenesis, inactive mutant Homo sapiens
additional information the CP1 hairpin of Homo sapiens cytoplasmic LeuRS (hcLeuRS) is deleted or substituted by those from other representative species. Lack of a CP1 hairpin leads to complete loss of aminoacylation, amino acid activation, and tRNA binding, butthe mutants retain post-transfer editing activity. Only the CP1 hairpin from Saccharomyces cerevisiae LeuRS (ScLeuRS) can partly rescue the hcLeuRS functions. Construction of chimeric mutants with the CP1 hairpin of hcLeuRS substituted for that of hcIleRS or hcValRS. The deacylating activity toward mischarged tRNALeu of hcLeuRS-ScCH1 and -ScCH2 decreases by 15% compared to that of hcLeuRS, kinetics comparisons, overview. Further site-directed mutagenesis indicates that the flexibility of small residues and the charge of polar residues in the CP1 hairpin are crucial for the function of LeuRS Homo sapiens
P242E site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
P247A site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
Q529A site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 70% reduced amino acid activation activity compared to the wild-type enzyme Homo sapiens
R236D site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 30% reduced amino acid activation activity compared to the wild-type enzyme Homo sapiens
R517D site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 90% reduced amino acid activation activity compared to the wild-type enzyme Homo sapiens
S519G site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
V523I site-directed mutagenesis, the mutant shows altered kinetics and reduced catalytic efficiency in the aminoacylation reaction compared to the wild-type enzyme Homo sapiens
W530A site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 50% reduced amino acid activation activity compared to the wild-type enzyme Homo sapiens
Y240A site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 50% reduced amino acid activation activity compared to the wild-type enzyme Homo sapiens
Y531A site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 50% reduced amino acid activation activity compared to the wild-type enzyme Homo sapiens
Y534A site-directed mutagenesis, the mutant shows altered kinetics, reduced catalytic efficiency in the aminoacylation reaction, and 60% reduced amino acid activation activity compared to the wild-type enzyme Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics of aminoacylation reaction of recombinant wild-type and mutant enzymes, and apparent kinetic parameters for hydrolytic editing of mischarged Met-tRNALeu, overview Homo sapiens

Localization

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

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-leucine + tRNALeu Homo sapiens
-
AMP + diphosphate + L-leucyl-tRNALeu
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens Q9P2J5
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-leucine + tRNALeu
-
Homo sapiens AMP + diphosphate + L-leucyl-tRNALeu
-
?

Subunits

Subunits Comment Organism
More enzyme LeuRS contains a catalysis domain (aminoacylation) and a CP1 domain (editing). CP1, residue R236 to G256, is inserted 35 A away from the aminoacylation domain. Aminoacylation and editing require CP1 to swing to the coordinated conformation. The neck between the CP1 domain and the aminoacylation domain is defined as the CP1 hairpin. The location of the CP1 hairpin suggests a crucial role in the CP1 swing and domain-domain interaction. Besides the conservative Rossmann fold, almost all LeuRSs contain a large insertion domain called connective peptide 1 (CP1) within the sequence of the catalytic core. CP1 folds independently in the tertiary structure and is defined as a classic editing domain, in which the aminoacyl bond of mischarged aatRNA is hydrolyzed (post-transfer editing) to ensure the fidelity of the catalytic process. Primary sequence and tertiary structure of the CP1 hairpin of LeuRS, and structure-function relationship, overview Homo sapiens

Synonyms

Synonyms Comment Organism
cytoplasmic LeuRS
-
Homo sapiens
HcleuRS
-
Homo sapiens
LARS
-
Homo sapiens
Leucyl-tRNA synthetase
-
Homo sapiens
LeuRS
-
Homo sapiens

Cofactor

Cofactor Comment Organism Structure
ATP
-
Homo sapiens

General Information

General Information Comment Organism
evolution leucyl-tRNA synthetase (LeuRS) belongs to class Ia aminoacyl-tRNA synthetases (AaRSs). Based on their similar structures, LeuRS, IleRS, and ValRS are collectively known as LIVRS, all of which contain a representative catalytic core consisting of a Rossmann fold. Besides the conservative Rossmann fold, almost all LeuRSs contain a large insertion domain called connective peptide 1 (CP1) within the sequence of the catalytic core. CP1 folds independently in the tertiary structure and is defined as a classic editing domain, in which the aminoacyl bond of mischarged aatRNA is hydrolyzed (post-transfer editing) to ensure the fidelity of the catalytic process Homo sapiens
additional information the CP1 hairpin editing structure, residue R236 to G256, and the flexibility of small residues and the charge of polar residues in the CP1 hairpin are crucial for the function of LeuRS. The CP1 hairpin domain is crucial for activities of leucine, leucylation of tRNALeu, and tRNA binding of hcLeuRS Homo sapiens
physiological function leucyl-tRNA synthetases (LeuRSs) catalyze the linkage of leucine with tRNALeu Homo sapiens