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Literature summary for 6.1.1.23 extracted from

  • Cardoso, A.M.; Polycarpo, C.; Martins, O.B.; Soell, D.
    A non-discriminating aspartyl-tRNA synthetase from Halobacterium salinarum (2006), RNA Biol., 3, 110-114.
    View publication on PubMed

Cloned(Commentary)

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

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

General Stability Organism
ND-AspRS is stable in low and high salt Halobacterium salinarum

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Halobacterium salinarum
NaCl required at 0.1-3 M, salt dependence profile, overview Halobacterium salinarum

Natural Substrates/ Products (Substrates)

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

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 (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant ND-AspRS in Escherichia coli strain trpA34 by nickel affinity chromatography Halobacterium salinarum

Substrates and Products (Substrate)

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

Synonyms Comment Organism
ND-AspRS
-
Halobacterium salinarum
non-discriminating aspartyl-tRNA synthetase
-
Halobacterium salinarum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Halobacterium salinarum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
aminoacylation assay at Halobacterium salinarum

Cofactor

Cofactor Comment Organism Structure
ATP
-
Halobacterium salinarum