Literature summary for 6.1.1.22 extracted from

Iwasaki, W.; Sekine, S.; Kuroishi, C.; Kuramitsu, S.; Shirouzu, M.; Yokoyama, S.
Structural basis of the water-assisted asparagine recognition by asparaginyl-tRNA synthetase (2006), J. Mol. Biol., 360, 329-342.

Cloned(Commentary)

Cloned (Comment)	Organism
overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Pyrococcus horikoshii

Crystallization (Commentary)

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

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

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

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

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Pyrococcus horikoshii

Organism

Organism	UniProt	Comment	Textmining
Pyrococcus horikoshii	O57980	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by anion exchange and hydroxyapatite chromatography	Pyrococcus horikoshii

Reaction

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	a

Substrates and Products (Substrate)

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

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 [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
60	-	assay at	Pyrococcus horikoshii

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Pyrococcus horikoshii