Cloned (Comment) | Organism |
---|---|
gene aspS, phylogenetic tree, expression of His-tagged wild-type and mutant ND-AspRS in Escherichia coli strain trpA34, carrying a D60N mutation in trpA leading to tryptophan auxotrophy, co-expression of tRNAAsn leading to restoration of tryptophan prototrophy by missense suppression of the trpA34 mutant with heterologously in vivo formed Asp-tRNAAsn | Halobacterium salinarum |
Protein Variants | Comment | Organism |
---|---|---|
H26A | site-directed mutagenesis, mutation the amino acid located in the AspRS anticodon binding domain limits the specificity of this nondiscriminating enzyme towards tRNAAsn, altered tRNA substrate specificity compared to the wild-type enzyme, overview | Halobacterium salinarum |
H26A/P84A | site-directed mutagenesis, mutation the amino acid located in the AspRS anticodon binding domain limits the specificity of this nondiscriminating enzyme towards tRNAAsn, altered tRNA substrate specificity compared to the wild-type enzyme, overview | Halobacterium salinarum |
H26A/P84K | site-directed mutagenesis, mutation the amino acid located in the AspRS anticodon binding domain limits the specificity of this nondiscriminating enzyme towards tRNAAsn, altered tRNA substrate specificity compared to the wild-type enzyme, overview | Halobacterium salinarum |
H26Q | site-directed mutagenesis, mutation the amino acid located in the AspRS anticodon binding domain limits the specificity of this nondiscriminating enzyme towards tRNAAsn, altered tRNA substrate specificity compared to the wild-type enzyme, overview | Halobacterium salinarum |
H26Q/P84A | site-directed mutagenesis, mutation the amino acid located in the AspRS anticodon binding domain limits the specificity of this nondiscriminating enzyme towards tRNAAsn, altered tRNA substrate specificity compared to the wild-type enzyme, overview | Halobacterium salinarum |
H26Q/P84K | site-directed mutagenesis, mutation the amino acid located in the AspRS anticodon binding domain limits the specificity of this nondiscriminating enzyme towards tRNAAsn, altered tRNA substrate specificity compared to the wild-type enzyme, overview | Halobacterium salinarum |
P84A | site-directed mutagenesis, mutation the amino acid located in the AspRS anticodon binding domain limits the specificity of this nondiscriminating enzyme towards tRNAAsn, altered tRNA substrate specificity compared to the wild-type enzyme, overview | Halobacterium salinarum |
P84K | site-directed mutagenesis, mutation the amino acid located in the AspRS anticodon binding domain limits the specificity of this nondiscriminating enzyme towards tRNAAsn, altered tRNA substrate specificity compared to the wild-type enzyme, overview | Halobacterium salinarum |
General Stability | Organism |
---|---|
ND-AspRS is stable in low and high salt | Halobacterium salinarum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Halobacterium salinarum | |
NaCl | required at 0.1-3 M, salt dependence profile, overview | Halobacterium salinarum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-aspartate + tRNAAsn | Halobacterium salinarum | reaction via transamidation mechanism | AMP + diphosphate + aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsn | Halobacterium salinarum NRC 1 | reaction via transamidation mechanism | AMP + diphosphate + aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsp | Halobacterium salinarum | - |
AMP + diphosphate + aspartyl-tRNAAsp | - |
? | |
ATP + L-aspartate + tRNAAsp | Halobacterium salinarum NRC 1 | - |
AMP + diphosphate + aspartyl-tRNAAsp | - |
? | |
additional information | Halobacterium salinarum | the tRNA-dependent transamidation pathway is the essential route for Asn-tRNAAsn formation in organisms that lack an asparaginyl-tRNA synthetase. This pathway relies on ND-AspRS, an enzyme with relaxed tRNA specificity, to form Asp-tRNAAsn, the misacylated tRNA is then converted to Asn-tRNAAsn by the action of an Asp-tRNAAsn amidotransferase, EC 6.3.5.6 | ? | - |
? | |
additional information | Halobacterium salinarum NRC 1 | the tRNA-dependent transamidation pathway is the essential route for Asn-tRNAAsn formation in organisms that lack an asparaginyl-tRNA synthetase. This pathway relies on ND-AspRS, an enzyme with relaxed tRNA specificity, to form Asp-tRNAAsn, the misacylated tRNA is then converted to Asn-tRNAAsn by the action of an Asp-tRNAAsn amidotransferase, EC 6.3.5.6 | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Halobacterium salinarum | O07683 | gene aspS, ND-AspRS; gene aspS | - |
Halobacterium salinarum NRC 1 | O07683 | gene aspS, ND-AspRS; gene aspS | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant ND-AspRS in Escherichia coli strain trpA34 by nickel affinity chromatography | Halobacterium salinarum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-aspartate + tRNAAsn | reaction via transamidation mechanism | Halobacterium salinarum | AMP + diphosphate + aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsn | in vivo expressed Halobacterium salinarum tRNAAsn, reaction via transamidation mechanism | Halobacterium salinarum | AMP + diphosphate + aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsn | reaction via transamidation mechanism | Halobacterium salinarum NRC 1 | AMP + diphosphate + aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsn | in vivo expressed Halobacterium salinarum tRNAAsn, reaction via transamidation mechanism | Halobacterium salinarum NRC 1 | AMP + diphosphate + aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsp | - |
Halobacterium salinarum | AMP + diphosphate + aspartyl-tRNAAsp | - |
? | |
ATP + L-aspartate + tRNAAsp | in vivo expressed Halobacterium salinarum tRNAAsp | Halobacterium salinarum | AMP + diphosphate + aspartyl-tRNAAsp | - |
? | |
ATP + L-aspartate + tRNAAsp | - |
Halobacterium salinarum NRC 1 | AMP + diphosphate + aspartyl-tRNAAsp | - |
? | |
additional information | the tRNA-dependent transamidation pathway is the essential route for Asn-tRNAAsn formation in organisms that lack an asparaginyl-tRNA synthetase. This pathway relies on ND-AspRS, an enzyme with relaxed tRNA specificity, to form Asp-tRNAAsn, the misacylated tRNA is then converted to Asn-tRNAAsn by the action of an Asp-tRNAAsn amidotransferase, EC 6.3.5.6 | Halobacterium salinarum | ? | - |
? | |
additional information | the Halobacterium salinarum enzyme is unable to use Escherichia coli tRNA as substrate | Halobacterium salinarum | ? | - |
? | |
additional information | the tRNA-dependent transamidation pathway is the essential route for Asn-tRNAAsn formation in organisms that lack an asparaginyl-tRNA synthetase. This pathway relies on ND-AspRS, an enzyme with relaxed tRNA specificity, to form Asp-tRNAAsn, the misacylated tRNA is then converted to Asn-tRNAAsn by the action of an Asp-tRNAAsn amidotransferase, EC 6.3.5.6 | Halobacterium salinarum NRC 1 | ? | - |
? | |
additional information | the Halobacterium salinarum enzyme is unable to use Escherichia coli tRNA as substrate | Halobacterium salinarum NRC 1 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ND-AspRS | - |
Halobacterium salinarum |
non-discriminating aspartyl-tRNA synthetase | - |
Halobacterium salinarum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Halobacterium salinarum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
aminoacylation assay at | Halobacterium salinarum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Halobacterium salinarum |