Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Pyrococcus horikoshii |
Crystallization (Comment) | Organism |
---|---|
AsnRS complexed with asparaginyl-adenylate Asn-AMP, or free AsnRS and AsnRS complexed with an Asn-AMP analog Asn-SA, sitting-drop vapor diffusion method at 20°C, 0.001 ml protein solution is mixed with an equal volume of reservoir solution containing 25 mM sodium cacodylate buffer, pH 6.5, 50 mM sodium acetate trihydrate, and 7.5% w/v PEG 8000, equilibration against 0.5 ml reservoir solution, for ligand complex formation the crystals are soaked in a reservoir solution, containing 1.5 mM AMP-PNP, 1.5 mM asparagine, and 2 mM MgCl2, for approximately 10 h, followed by a reservoir solution containing 25% 2-methyl-2,4-pentanediol for a few seconds, X-ray diffraction strucure determination and analysis at 1.45 A, and 1.98 A and 1.8 A resolution, respectively | Pyrococcus horikoshii |
Protein Variants | Comment | Organism |
---|---|---|
R83A | site-directed mutagenesis, active site amino acid binding mutant, structure analysis in comparison to the wild-type enzyme | Pyrococcus horikoshii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5'-O-[N-(L-asparaginyl)sulfamoyl]adenosine | a non-hydrolysable analogue of asparaginyl adenylate, i.e. Asn-SA | Pyrococcus horikoshii |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Pyrococcus horikoshii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus horikoshii | O57980 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by anion exchange and hydroxyapatite chromatography | Pyrococcus horikoshii |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + L-asparagine + tRNAAsn = AMP + diphosphate + L-asparaginyl-tRNAAsn | substrate binding structure, active site structure, tRNAAsn and amino acid recognition, binding of Asn-AMP induces a conformational change in AsnRS | Pyrococcus horikoshii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme is responsible for catalyzing the specific aminoacylation of tRNAAsn with asparagine, structural basis of the water-assisted asparagine recognition by the enzyme, two water molecules play a key role in the strict recognition of asparagine and the discrimination against aspartic acid by the AsnRS, overview, binding of Asn-AMP induces a conformational change in AsnRS | Pyrococcus horikoshii | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AsnRS | - |
Pyrococcus horikoshii |
Asparaginyl-tRNA synthetase | - |
Pyrococcus horikoshii |
More | the enzyme belongs to the class-II aminoacyl-tRNA synthetase family | Pyrococcus horikoshii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
assay at | Pyrococcus horikoshii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Pyrococcus horikoshii |