Cloned (Comment) | Organism |
---|---|
gene CDC60, expression of wild-type and mutants in an Escherichia coli BL21 strain | Saccharomyces cerevisiae |
gene leuS, expression in a BL21 strain | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
crystal growth in presence of mercuric chloride, soaking of the crystals in solution containing 0.6 mM of the non-hydrolyzable substrate analogue norvaline-AMS for 1 month, or cocrystallization of enzyme and norvaline-AMS, X-ray diffraction structure determinationat 2.0-2.2 A resolution and analysis | Thermus thermophilus |
Protein Variants | Comment | Organism |
---|---|---|
D345A | mutation of the highly conserved Asp residue, located in the CP1 domain, is responsible for editing mechanism, slightly reduced activity with L-leucine, mutant mischarges tRNALeu with isoleucine | Escherichia coli |
D347A | mutation of the highly conserved Asp residue, located in the CP1 domain, is responsible for editing mechanism, slightly reduced activity with L-leucine, mutant mischarges tRNALeu with isoleucine | Thermus thermophilus |
D419A | mutation of the highly conserved Asp residue, located in the CP1 domain, is responsible for editing mechanism, slightly reduced activity with L-leucine, mutant mischarges tRNALeu with isoleucine | Saccharomyces cerevisiae |
T252A | decreased activity with L-leucine, mutant shows altered editing specificity, it edits correctly formed leucyl-tRNALeu | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
O-[N-(L-norvalyl)sulfamoyl]adenosine | analogue to the reaction intermediate, non-hydrolyzable | Thermus thermophilus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytoplasm | - |
Saccharomyces cerevisiae | 5737 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-leucine + tRNALeu | Escherichia coli | - |
AMP + diphosphate + L-leucyl-tRNALeu | - |
r | |
ATP + L-leucine + tRNALeu | Saccharomyces cerevisiae | - |
AMP + diphosphate + L-leucyl-tRNALeu | - |
r | |
ATP + L-leucine + tRNALeu | Thermus thermophilus | - |
AMP + diphosphate + L-leucyl-tRNALeu | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Saccharomyces cerevisiae | - |
- |
- |
Thermus thermophilus | Q72GM3 | purified recombinant enzyme | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | Escherichia coli |
recombinant from Escherichia coli | Saccharomyces cerevisiae |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu | active site structure and mechanism, the editing active site binds the two different substrates using a single amino acid discriminatory pocket while preserving he same mode of adenine recognition, Asp347 is involved in the editing process, mechanism of hydrolysis | Thermus thermophilus | |
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu | Asp345 is involved in the editing process, mechanism of hydrolysis | Escherichia coli | |
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu | Asp419 is involved in the editing process, mechanism of hydrolysis | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-isoleucine + tRNALeu | mutant D345A, not the wild-type which performs only the misacetylation with isoleucine, but eliminates the incorrect isoleucyl-AMP | Escherichia coli | AMP + diphosphate + L-isoleucyl-tRNALeu | - |
r | |
ATP + L-isoleucine + tRNALeu | mutant D345A, not the wild-type which performs only the misacetylation with isoleucine, but eliminates the incorrect isoleucyl-AMP | Thermus thermophilus | AMP + diphosphate + L-isoleucyl-tRNALeu | - |
r | |
ATP + L-isoleucine + tRNALeu | mutant D419A, not the wild-type, which performs only the misacetylation with isoleucine, but eliminates the incorrect isoleucyl-AMP | Saccharomyces cerevisiae | AMP + diphosphate + L-isoleucyl-tRNALeu | - |
r | |
ATP + L-leucine + tRNALeu | - |
Escherichia coli | AMP + diphosphate + L-leucyl-tRNALeu | - |
r | |
ATP + L-leucine + tRNALeu | - |
Saccharomyces cerevisiae | AMP + diphosphate + L-leucyl-tRNALeu | - |
r | |
ATP + L-leucine + tRNALeu | - |
Thermus thermophilus | AMP + diphosphate + L-leucyl-tRNALeu | - |
r | |
ATP + L-leucine + tRNALeu | mutant T252A edits correctly charged Leu-tRNALeu | Escherichia coli | AMP + diphosphate + L-leucyl-tRNALeu | - |
r | |
ATP + L-leucine + tRNALeu | the editing active site hydrolytically cleaves the misactivated aminoacyl-adenylate, called pre-transfer editing, or the mischarged tRNA, called post-transfer editing | Thermus thermophilus | AMP + diphosphate + L-leucyl-tRNALeu | - |
r | |
ATP + L-methionine + tRNALeu | mutant D345A, not the wild-type enzyme | Escherichia coli | AMP + diphosphate + L-methionyl-tRNALeu | - |
r | |
ATP + L-methionine + tRNALeu | mutant D419A, not the wild-type enzyme | Saccharomyces cerevisiae | AMP + diphosphate + L-methionyl-tRNALeu | - |
r |
Synonyms | Comment | Organism |
---|---|---|
Leucine translase | - |
Escherichia coli |
Leucine translase | - |
Saccharomyces cerevisiae |
Leucine translase | - |
Thermus thermophilus |
Leucine--tRNA ligase | - |
Escherichia coli |
Leucine--tRNA ligase | - |
Saccharomyces cerevisiae |
Leucine--tRNA ligase | - |
Thermus thermophilus |
Leucyl-transfer ribonucleate synthetase | - |
Escherichia coli |
Leucyl-transfer ribonucleate synthetase | - |
Saccharomyces cerevisiae |
Leucyl-transfer ribonucleate synthetase | - |
Thermus thermophilus |
Leucyl-transfer ribonucleic acid synthetase | - |
Escherichia coli |
Leucyl-transfer ribonucleic acid synthetase | - |
Saccharomyces cerevisiae |
Leucyl-transfer ribonucleic acid synthetase | - |
Thermus thermophilus |
Leucyl-transfer RNA synthetase | - |
Escherichia coli |
Leucyl-transfer RNA synthetase | - |
Saccharomyces cerevisiae |
Leucyl-transfer RNA synthetase | - |
Thermus thermophilus |
Leucyl-tRNA synthetase | - |
Escherichia coli |
Leucyl-tRNA synthetase | - |
Saccharomyces cerevisiae |
Leucyl-tRNA synthetase | - |
Thermus thermophilus |
LeuRS | - |
Escherichia coli |
LeuRS | - |
Saccharomyces cerevisiae |
LeuRS | - |
Thermus thermophilus |
LeuRSTT | - |
Thermus thermophilus |
Synthetase, leucyl-transfer ribonucleate | - |
Escherichia coli |
Synthetase, leucyl-transfer ribonucleate | - |
Saccharomyces cerevisiae |
Synthetase, leucyl-transfer ribonucleate | - |
Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
7.5 | - |
assay at | Saccharomyces cerevisiae |
7.5 | - |
assay at | Thermus thermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli | |
ATP | - |
Saccharomyces cerevisiae | |
ATP | - |
Thermus thermophilus |