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Literature summary for 6.1.1.4 extracted from

  • Hu, Q.H.; Huang, Q.; Wang, E.D.
    Crucial role of the C-terminal domain of Mycobacterium tuberculosis leucyl-tRNA synthetase in aminoacylation and editing (2013), Nucleic Acids Res., 41, 1859-1872.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
L949A the mutant shows 9.5% activity compared to the wild type enzyme Mycobacterium tuberculosis
L949K the mutant shows 2.6% activity compared to the wild type enzyme Mycobacterium tuberculosis
L964A the mutant shows 85% activity compared to the wild type enzyme Mycobacterium tuberculosis
L964K the mutant shows 8.6% activity compared to the wild type enzyme Mycobacterium tuberculosis
Q915A the mutant shows 99% activity compared to the wild type enzyme Mycobacterium tuberculosis
Q915K the mutant shows 54% activity compared to the wild type enzyme Mycobacterium tuberculosis
R921A the mutant shows 37% activity compared to the wild type enzyme Mycobacterium tuberculosis
R921K the mutant shows 83% activity compared to the wild type enzyme Mycobacterium tuberculosis
V910A the mutant shows 90% activity compared to the wild type enzyme Mycobacterium tuberculosis
V910P the mutant shows 3.7% activity compared to the wild type enzyme Mycobacterium tuberculosis
V910W the mutant shows 93% activity compared to the wild type enzyme Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0011
-
tRNALeu mutant enzyme V910A, at pH 8.2 and 30°C Mycobacterium tuberculosis
0.0011
-
tRNALeu wild type enzyme, at pH 8.2 and 30°C Mycobacterium tuberculosis
0.0012
-
tRNALeu mutant enzyme L964A, at pH 8.2 and 30°C Mycobacterium tuberculosis
0.0013
-
tRNALeu mutant enzyme Q915K, at pH 8.2 and 30°C Mycobacterium tuberculosis
0.0014
-
tRNALeu mutant enzyme V910W, at pH 8.2 and 30°C Mycobacterium tuberculosis
0.0018
-
tRNALeu mutant enzyme R921K, at pH 8.2 and 30°C Mycobacterium tuberculosis
0.0019
-
tRNALeu mutant enzyme Q915A, at pH 8.2 and 30°C Mycobacterium tuberculosis
0.0052
-
tRNALeu mutant enzyme R921A, at pH 8.2 and 30°C Mycobacterium tuberculosis
0.0083
-
tRNALeu mutant enzyme L949A, at pH 8.2 and 30°C Mycobacterium tuberculosis
0.0092
-
tRNALeu mutant enzyme V910P, at pH 8.2 and 30°C Mycobacterium tuberculosis
0.0174
-
tRNALeu mutant enzyme L964K, at pH 8.2 and 30°C Mycobacterium tuberculosis
0.0273
-
tRNALeu mutant enzyme L949K, at pH 8.2 and 30°C Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ the optimum Mg2+:ATP ratio for aminoacylation is 3:1 Mycobacterium tuberculosis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
108000
-
x * 108000, calculated from amino acid sequence Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-leucine + tRNALeu Mycobacterium tuberculosis
-
AMP + diphosphate + L-leucyl-tRNALeu
-
?
ATP + L-leucine + tRNALeu Mycobacterium tuberculosis H37Rv
-
AMP + diphosphate + L-leucyl-tRNALeu
-
?

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
-
-
Mycobacterium tuberculosis H37Rv
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni2+-NTA column chromatography Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-leucine + tRNALeu
-
Mycobacterium tuberculosis AMP + diphosphate + L-leucyl-tRNALeu
-
?
ATP + L-leucine + tRNALeu
-
Mycobacterium tuberculosis H37Rv AMP + diphosphate + L-leucyl-tRNALeu
-
?

Subunits

Subunits Comment Organism
? x * 108000, calculated from amino acid sequence Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
Leucyl-tRNA synthetase
-
Mycobacterium tuberculosis
LeuRS
-
Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
-
Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4
-
tRNALeu mutant enzyme V910P, at pH 8.2 and 30°C Mycobacterium tuberculosis
5
-
tRNALeu mutant enzyme L949K, at pH 8.2 and 30°C Mycobacterium tuberculosis
5
-
tRNALeu mutant enzyme Q915K, at pH 8.2 and 30°C Mycobacterium tuberculosis
5.6
-
tRNALeu mutant enzyme L949A, at pH 8.2 and 30°C Mycobacterium tuberculosis
7
-
tRNALeu mutant enzyme V910A, at pH 8.2 and 30°C Mycobacterium tuberculosis
7.2
-
tRNALeu mutant enzyme L964A, at pH 8.2 and 30°C Mycobacterium tuberculosis
7.8
-
tRNALeu wild type enzyme, at pH 8.2 and 30°C Mycobacterium tuberculosis
9.2
-
tRNALeu mutant enzyme V910W, at pH 8.2 and 30°C Mycobacterium tuberculosis
10.6
-
tRNALeu mutant enzyme L964K, at pH 8.2 and 30°C Mycobacterium tuberculosis
10.6
-
tRNALeu mutant enzyme R921K, at pH 8.2 and 30°C Mycobacterium tuberculosis
13.3
-
tRNALeu mutant enzyme Q915A, at pH 8.2 and 30°C Mycobacterium tuberculosis
13.6
-
tRNALeu mutant enzyme R921A, at pH 8.2 and 30°C Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.2
-
-
Mycobacterium tuberculosis

Cofactor

Cofactor Comment Organism Structure
ATP
-
Mycobacterium tuberculosis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
183.2
-
tRNALeu mutant enzyme L949K, at pH 8.2 and 30°C Mycobacterium tuberculosis
260.9
-
tRNALeu mutant enzyme V910P, at pH 8.2 and 30°C Mycobacterium tuberculosis
609.2
-
tRNALeu mutant enzyme L964K, at pH 8.2 and 30°C Mycobacterium tuberculosis
674.7
-
tRNALeu mutant enzyme L949A, at pH 8.2 and 30°C Mycobacterium tuberculosis
2615
-
tRNALeu mutant enzyme R921A, at pH 8.2 and 30°C Mycobacterium tuberculosis
3846
-
tRNALeu mutant enzyme Q915K, at pH 8.2 and 30°C Mycobacterium tuberculosis
5889
-
tRNALeu mutant enzyme R921K, at pH 8.2 and 30°C Mycobacterium tuberculosis
6000
-
tRNALeu mutant enzyme L964A, at pH 8.2 and 30°C Mycobacterium tuberculosis
6364
-
tRNALeu mutant enzyme V910A, at pH 8.2 and 30°C Mycobacterium tuberculosis
6571
-
tRNALeu mutant enzyme V910W, at pH 8.2 and 30°C Mycobacterium tuberculosis
7000
-
tRNALeu mutant enzyme Q915A, at pH 8.2 and 30°C Mycobacterium tuberculosis
7091
-
tRNALeu wild type enzyme, at pH 8.2 and 30°C Mycobacterium tuberculosis