Literature summary for 6.1.1.24 extracted from

Ito, T.; Kiyasu, N.; Matsunaga, R.; Takahashi, S.; Yokoyama, S.
Structure of nondiscriminating glutamyl-tRNA synthetase from Thermotoga maritima (2010), Acta Crystallogr. Sect. D, 66, 813-820.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli	Thermotoga maritima
TM1875, sequence comparisons, expression in Escherichia coli strain Rosetta2(DE3)	Thermotoga maritima

Crystallization (Commentary)

Crystallization (Comment)	Organism
the crystal structure of the Thermotoga maritima ND-GluRS, TM1875, is determined in complex with a Glu-AMP analogue at 2.0 A resolution. ND-GluRS contains a characteristic structure in the connective-peptide domain, which is inserted into the catalytic Rossmann-fold domain	Thermotoga maritima
TM1875 in complex with a Glu-AMP analogue, mixing of 0.0018 ml of 15 mg/ml protein in 20 mM Tris-HCl, pH 7.0, containing 5 mM MgCl2, 10 mM 2-mercaptoethanol, 50 mM NaCl, and 1 mM L-glutamylsulfamoyl adenosine, with 0.0018 ml reservoir solution containing 100 mM HEPES-NaOH, pH 7.5, 8% ethylene glycol and 15% PEG 8000, and 0.0004 ml of 30% D-sorbitol, equilibration against 0.5 ml reservoir solution, 20°C, 1 week, X-ray diffraction structure determination and analysis at 2.0 A resolution	Thermotoga maritima

Crystallization (Comment)

Organism

the crystal structure of the Thermotoga maritima ND-GluRS, TM1875, is determined in complex with a Glu-AMP analogue at 2.0 A resolution. ND-GluRS contains a characteristic structure in the connective-peptide domain, which is inserted into the catalytic Rossmann-fold domain

Thermotoga maritima

TM1875 in complex with a Glu-AMP analogue, mixing of 0.0018 ml of 15 mg/ml protein in 20 mM Tris-HCl, pH 7.0, containing 5 mM MgCl2, 10 mM 2-mercaptoethanol, 50 mM NaCl, and 1 mM L-glutamylsulfamoyl adenosine, with 0.0018 ml reservoir solution containing 100 mM HEPES-NaOH, pH 7.5, 8% ethylene glycol and 15% PEG 8000, and 0.0004 ml of 30% D-sorbitol, equilibration against 0.5 ml reservoir solution, 20°C, 1 week, X-ray diffraction structure determination and analysis at 2.0 A resolution

Thermotoga maritima

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Thermotoga maritima
Mg2+	required	Thermotoga maritima

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
additional information	Thermotoga maritima	ND-GluRS TM1875 glutamylates both tRNAGlu and tRNAGln	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Thermotoga maritima	Q9X2I8	-	-
Thermotoga maritima	Q9X2I8	TM1875	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant TM1875 from Escherichia coli strain Rosetta2 (DE3) by anion exchange chromatography and gel filtration	Thermotoga maritima
using ion-exchange column chromatography and gel-filtration	Thermotoga maritima

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + Glu + tRNAGln	the glutamylation ability of tRNAGln by ND-GluRS is measured in the presence of the bacterial Glu-tRNAGln amidotransferase GatCAB. Glutamylation efficiency is not affected even in the presence of excess GatCAB	Thermotoga maritima	AMP + diphosphate + L-glutamyl-tRNAGln	-	?
additional information	ND-GluRS TM1875 glutamylates both tRNAGlu and tRNAGln	Thermotoga maritima	?	-	?
additional information	TM1875 glutamylates both the tRNAGlu and tRNAGln transcripts	Thermotoga maritima	?	-	?

Subunits

Subunits	Comment	Organism
More	Thermotoga maritima ND-GluRS contains a characteristic structure in the connective-peptide domain, which is inserted into the catalytic Rossmann-fold domain, structure, overview	Thermotoga maritima

Synonyms

Synonyms	Comment	Organism
Glutamyl-tRNA synthetase	-	Thermotoga maritima
ND-GluRS	-	Thermotoga maritima
nondiscriminating GluRS	-	Thermotoga maritima
nondiscriminating glutamyl-tRNA synthetase	-	Thermotoga maritima
TM1875	-	Thermotoga maritima
TM1875	only TM1875 functions as a nondiscriminating GluRS	Thermotoga maritima

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
50	-	assay at	Thermotoga maritima

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Thermotoga maritima

General Information

General Information	Comment	Organism
physiological function	ND-GluRS produces the intermediate Glu-tRNAGln, which is converted to Gln-tRNAGln by Glu-tRNAGln amidotransferase. GluRS avoids competition with Glu-tRNAGln amidotransferase GatCAB and glutamylates tRNAGln	Thermotoga maritima