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Literature summary for 6.3.5.6 extracted from
- The heterotrimeric Thermus thermophilus Asp-tRNA(Asn) amidotransferase can also generate Gln-tRNA(Gln) (2000), FEBS Lett., 476, 140-144.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| canonical set of amidotransferase genes gatA, gatB and gatC is cloned and sequenced, the holoenzyme is overexpessed in Escherichia coli and exhibits both Asp-tRNAAsn and Glu-tRNAGln transamidation activities, the gatA, gatB and gatC genes are dispersed in the genome and encode polypeptides of 472, 470 and 90 amino acids, removal of the 44 carboxy-terminal amino acids of the GatA subunit only inhibits the Asp-AdT activity, but not the Glu-AdT activity | Thermus thermophilus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | GatDDELTA44 mutant AdT possesses a GatA subunit deprived of its natural 44 C-terminal amino acids and shows unaltered Glu-AdT activity, but lacks any detectable Asp-AdT activity, deletion of one base in comparison with the HB27 genome causes a frameshift mutation | Thermus thermophilus |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| Asp-tRNAAsn + ? | Thermus thermophilus | in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity | Asn-tRNAAsn + ? | - |
? |  |
| Asp-tRNAAsn + ? | Deinococcus radiodurans | in vivo only Asn-tRNAAsn formation occurs | Asn-tRNAAsn + ? | - |
? | |
| Asp-tRNAAsn + ? | Thermus thermophilus | enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase | Asn-tRNAAsn + ? | - |
? | |
| Asp-tRNAAsn + ? | Deinococcus radiodurans | enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase | Asn-tRNAAsn + ? | - |
? | |
| Asp-tRNAAsn + ? | Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity | Asn-tRNAAsn + ? | - |
? | |
| Asp-tRNAAsn + ? | Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase | Asn-tRNAAsn + ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus subtilis | - |
- |
- |
| Deinococcus radiodurans | - |
- |
- |
| Thermus thermophilus | - |
and strain HB27 / ATCC BAA-163 / DSM 7039 | - |
| Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | - |
and strain HB27 / ATCC BAA-163 / DSM 7039 | - |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| Asp-tRNAAsn + ? | in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity | Thermus thermophilus | Asn-tRNAAsn + ? | - |
? | |
| Asp-tRNAAsn + ? | in vivo only Asn-tRNAAsn formation occurs | Deinococcus radiodurans | Asn-tRNAAsn + ? | - |
? | |
| Asp-tRNAAsn + ? | enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase | Thermus thermophilus | Asn-tRNAAsn + ? | - |
? | |
| Asp-tRNAAsn + ? | enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase | Deinococcus radiodurans | Asn-tRNAAsn + ? | - |
? | |
| Asp-tRNAAsn + ? | in vivo only Asn-tRNAAsn formation occurs, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, and cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases, explaining why those enzymes have not restricted their specificity | Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | Asn-tRNAAsn + ? | - |
? | |
| Asp-tRNAAsn + ? | enzyme of the two-step pathway for the synthesis of Asn-tRNA that requires misacylated Asp-tRNAAsn as intermediate, which is generated by a Asp-tRNA synthetase | Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | Asn-tRNAAsn + ? | - |
? | |
| ATP + Asp-tRNAAsn + glutamine | - |
Bacillus subtilis | ADP + phosphate + Asn-tRNAAsn + glutamate | - |
? | |
| ATP + Asp-tRNAAsn + glutamine | - |
Deinococcus radiodurans | ADP + phosphate + Asn-tRNAAsn + glutamate | - |
? | |
| ATP + Asp-tRNAAsn + glutamine | no restricted substrate specificity of tRNA-dependent AdT: dual-specific Asp/Glu-AdT, enzyme catalyzes transamidation of Asp-tRNAAsn, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, GatA subunit can discriminate between Asp-tRNAAsn and Glu-tRNAGln and therefore is implicated in binding of the tRNA | Thermus thermophilus | ADP + phosphate + Asn-tRNAAsn + glutamate | forms Asn-tRNAAsn | ? | |
| ATP + Asp-tRNAAsn + glutamine | no restricted substrate specificity of tRNA-dependent AdT: dual-specific Asp/Glu-AdT, enzyme catalyzes transamidation of Asp-tRNAAsn, since Thermus possesses a mischarging Asp-tRNA synthetase, but lacks a mischarging Glu-tRNA synthetase, GatA subunit can discriminate between Asp-tRNAAsn and Glu-tRNAGln and therefore is implicated in binding of the tRNA | Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | ADP + phosphate + Asn-tRNAAsn + glutamate | forms Asn-tRNAAsn | ? | |
| ATP + Glu-tRNAGln + glutamine | in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Gln-tRNAGln formation | Bacillus subtilis | ADP + Gln-tRNAGln + glutamate + phosphate | - |
? | |
| ATP + Glu-tRNAGln + glutamine | in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Asn-tRNAAsn formation | Thermus thermophilus | ADP + Gln-tRNAGln + glutamate + phosphate | - |
? | |
| ATP + Glu-tRNAGln + glutamine | in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Asn-tRNAAsn formation | Deinococcus radiodurans | ADP + Gln-tRNAGln + glutamate + phosphate | - |
? | |
| ATP + Glu-tRNAGln + glutamine | in vitro both Asp-tRNAAsn and Glu-tRNAGln amidation activities, in vivo only Asn-tRNAAsn formation | Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | ADP + Gln-tRNAGln + glutamate + phosphate | - |
? | |
| additional information | - |
Deinococcus radiodurans | ? | - |
? | |
| additional information | cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases | Thermus thermophilus | ? | - |
? | |
| additional information | cellular activity of AdTs only depends upon presence or lack of the mischarging amino acyl-tRNA synthetases | Thermus thermophilus HB8 / ATCC 27634 / DSM 579 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| heterodimer | Asp-AdT is active as a GatAB heterodimer | Thermus thermophilus |
| heterotrimer | - |
Bacillus subtilis |
| heterotrimer | - |
Deinococcus radiodurans |
| heterotrimer | possesses the three canonical gatA, gatB and gatC set of genes encoding heterotrimeric AdT: GatABC, GatA and GatB are more tightly bound to each other than to GatC, GatA subunit has conserved Gln-436 | Thermus thermophilus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at, suboptimal temperature for the thermophilic enzyme | Thermus thermophilus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
thermophilic enzyme | Thermus thermophilus |
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