Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in yeast YBC-603 cells, functional overexpression of GST- and His6-tagged enzyme in Escherichia coli strain Top10. Expression in yeast cells at low copy number, since overexpression is cytotoxic, while moderate AspRS accumulation in the cell is not deleterious | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
additional information | proteomic analysis of mutant cells in comparison to wild-type cells | Saccharomyces cerevisiae |
R485K | site-directed mutagenesis, the substitution in the catalytic site completely inhibits aspartylation by impairing ATP binding, this mutant still retains the capacity to be modified and shows the same pattern as wild-type AspRS on the two-dimensional gel, thus the modifications are not the result of autoaspartylation | Saccharomyces cerevisiae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | the yeast AspRS initiates retro-inhibition of its expression, overview | Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.018 | - |
tRNAAsp | - |
Saccharomyces cerevisiae |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Saccharomyces cerevisiae | 5829 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-aspartate + tRNAAsp | Saccharomyces cerevisiae | the yeast AspRS not only binds and aminoacylates tRNAAsp but also binds its yeast mRNA and initiates retro-inhibition of its expression, overview | AMP + diphosphate + L-aspartyl-tRNAAsp | - |
? | |
additional information | Saccharomyces cerevisiae | misaspartylation of tRNAAsn and tRNAGlu does not exist in vivo | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
additional information | the enzyme does not perform autoaspartylation in vivo | Saccharomyces cerevisiae |
no glycoprotein | - |
Saccharomyces cerevisiae |
Purification (Comment) | Organism |
---|---|
recombinant GST-tagged enzyme from Escherichia coli strain Top10 by glutathione affinity chromatography, the tag is then cleaved of by thrombin, and the enzyme is further purified by adsorption chromatography yielding an AspRS with a short additional amino acid stretch at its N-terminus | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-aspartate + tRNAAsp | the yeast AspRS not only binds and aminoacylates tRNAAsp but also binds its yeast mRNA and initiates retro-inhibition of its expression, overview | Saccharomyces cerevisiae | AMP + diphosphate + L-aspartyl-tRNAAsp | - |
? | |
ATP + L-aspartate + tRNAAsp | the yeast enzyme also binds its own mRNA and autoinhibits itself | Saccharomyces cerevisiae | AMP + diphosphate + L-aspartyl-tRNAAsp | - |
? | |
additional information | misaspartylation of tRNAAsn and tRNAGlu does not exist in vivo | Saccharomyces cerevisiae | ? | - |
? | |
additional information | increased concentrations of AspRS lead to the accumulation of significant amounts of Asp-tRNAAsn and Asp-tRNAGlu in vitro, but not in vivo. The enzyme does not perform autoaspartylation | Saccharomyces cerevisiae | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Aspartyl-tRNA synthetase | - |
Saccharomyces cerevisiae |
AspRS | - |
Saccharomyces cerevisiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Saccharomyces cerevisiae |