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Literature summary for 6.1.1.4 extracted from

  • Zhou, X.L.; Yao, P.; Ruan, L.L.; Zhu, B.; Luo, J.; Qu, L.H.; Wang, E.D.
    A unique insertion in the CP1 domain of Giardia lamblia leucyl-tRNA synthetase (2009), Biochemistry, 48, 1340-1347.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Giardia intestinalis

Protein Variants

Protein Variants Comment Organism
E298A activity similar to wild-type Giardia intestinalis
K299A activity similar to wild-type Giardia intestinalis
K303A activity similar to wild-type Giardia intestinalis
additional information construction of deletion mutant LeuRS-A lacking residues Q281 to D294, and of deletion mutant LeuRS-B, lacking residues S295 to L304. The Km values of the two mutants for leucine and ATP decrease slightly and for tRNA increase slightly. The kcat values for the three substrates decrease Giardia intestinalis
N301A activity similar to wild-type Giardia intestinalis
S295A activity similar to wild-type Giardia intestinalis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0059
-
L-leucine mutant lacking residues Q281 to D294, 45°C Giardia intestinalis
0.0061
-
tRNALeu wild-type, 45°C Giardia intestinalis
0.0077
-
L-leucine mutant lacking residues S295 to L304, 45°C Giardia intestinalis
0.0082
-
tRNALeu mutant lacking residues S295 to L304, 45°C Giardia intestinalis
0.009
-
L-leucine wild-type, 45°C Giardia intestinalis
0.0101
-
tRNALeu mutant lacking residues Q281 to D294, 45°C Giardia intestinalis
0.33
-
ATP mutant lacking residues Q281 to D294, 45°C Giardia intestinalis
0.688
-
ATP wild-type, 45°C Giardia intestinalis

Organism

Organism UniProt Comment Textmining
Giardia intestinalis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant protein Giardia intestinalis

Substrates and Products (Substrate)

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

Subunits

Subunits Comment Organism
? the isolated CP1 domain of LeuRS is an active protein for editing mischarged tRNALeu(AAG). An insertion of 49 amino acid residues within the CP1 domain, the so-called 49-amino acid motif, is important for the optimal aminoacylation activity of LeuRS and is crucial for the editing capacity of LeuRS-CP1 Giardia intestinalis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7
-
tRNALeu mutant lacking residues Q281 to D294, 45°C Giardia intestinalis
2.7
-
tRNALeu mutant lacking residues S295 to L304, 45°C Giardia intestinalis
2.7
-
tRNALeu wild-type, 45°C Giardia intestinalis
2.8
-
L-leucine mutant lacking residues Q281 to D294, 45°C Giardia intestinalis
2.8
-
L-leucine mutant lacking residues S295 to L304, 45°C Giardia intestinalis
2.8
-
L-leucine wild-type, 45°C Giardia intestinalis
3.1
-
ATP mutant lacking residues Q281 to D294, 45°C Giardia intestinalis
3.1
-
ATP mutant lacking residues S295 to L304, 45°C Giardia intestinalis
3.1
-
ATP wild-type, 45°C Giardia intestinalis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.2
-
ATP mutant lacking residues S295 to L304, 45°C Giardia intestinalis
3.6
-
ATP mutant lacking residues Q281 to D294, 45°C Giardia intestinalis
4.5
-
ATP wild-type, 45°C Giardia intestinalis
73.2
-
tRNALeu mutant lacking residues S295 to L304, 45°C Giardia intestinalis
77.9
-
L-leucine mutant lacking residues S295 to L304, 45°C Giardia intestinalis
128.7
-
tRNALeu mutant lacking residues Q281 to D294, 45°C Giardia intestinalis
186
-
L-leucine mutant lacking residues Q281 to D294, 45°C Giardia intestinalis
311
-
L-leucine wild-type, 45°C Giardia intestinalis
442
-
tRNALeu wild-type, 45°C Giardia intestinalis