Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Giardia intestinalis |
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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Giardia intestinalis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant protein | Giardia intestinalis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-leucine + tRNALeu | - |
Giardia intestinalis | AMP + diphosphate + L-leucyl-tRNALeu | - |
? |
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 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 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 |