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Literature summary for 6.1.1.24 extracted from

  • Bullock, T.L.; Rodriguez-Hernandez, A.; Corigliano, E.M.; Perona, J.J.
    A rationally engineered misacylating aminoacyl-tRNA synthetase (2008), Proc. Natl. Acad. Sci. USA, 105, 7428-7433.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

Crystallization (Comment) Organism
GlnRS-tRNAGln complex, 6.6 mg/ml protein in 10 mM PIPES, pH 7.5, 10 mM MgCl2, and 1.8-5.4 mM tRNA. The tRNA/analog solution is then mixed with equal volumes of a 6.3 mg/ml solution of GlnRS, containing 5mM PIPES, pH 7.0, and 5 mM 2-mercaptoethanol, X-ray diffraction structure determination and analysis at 2.6 A resolution Escherichia coli

Protein Variants

Protein Variants Comment Organism
additional information the engineered mutant hybrid C229R Gln-RS, EC 6.1.1.18, shows activity with L-glutamine or L-glutamate and tRNAGln like the nondiscriminating enzyme, EC 6.1.1.24. Introduction of 22 amino acid replacements and one deletion, including substitution of the entire primary binding site and two surface loops adjacent to the region disrupted in the mutant C229R, improves the capacity of the mutant enzyme to synthesize misacylated Glu-tRNAGln by 16000fold, overview Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics of the mutant enzyme compared to wild-type GlnRS, EC 6.1.1.18, overview Escherichia coli
0.21
-
L-glutamine mutant C229R GlnRS, with tRNAGln Escherichia coli
240
-
L-glutamate mutant C229R GlnRS, with tRNAGln Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-glutamate + tRNAGln Escherichia coli
-
AMP + diphosphate + L-glutamyl-tRNAGln
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli P00962
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-glutamate + tRNAGln
-
Escherichia coli AMP + diphosphate + L-glutamyl-tRNAGln
-
?
ATP + L-glutamate + tRNAGln activity also with Gln-RS, EC 6.1.1.18, mutant C229R Escherichia coli AMP + diphosphate + L-glutamyl-tRNAGln
-
?
ATP + L-glutamine + tRNAGln
-
Escherichia coli AMP + diphosphate + glutaminyl-tRNAGln
-
?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
assay at room temperature Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.00041
-
L-glutamate mutant C229R GlnRS, with tRNAGln Escherichia coli
0.0025
-
L-glutamine mutant C229R GlnRS, with tRNAGln Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
Escherichia coli