Literature summary for 6.1.1.18 extracted from

Ognjenovic, J.; Wu, J.; Matthies, D.; Baxa, U.; Subramaniam, S.; Ling, J.; Simonovic, M.
The crystal structure of human GlnRS provides basis for the development of neurological disorders (2016), Nucleic Acids Res., 44, 3420-3431 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene QARS, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta (DE3)pLysS	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged wild-type and mutant enzymes, sitting drop vapour diffusion method, apo wild-type GlnRS crystallizes from 0.1 M calcium acetate, 0.1 M Tris, pH 6.0, 12.5% w/v PEG 3350, and 60 mM Gly-Gly-Gly, the Y57H mutant crystallizes from 0.1 M ammonium acetate, 0.1 M Bis-Tris, pH 5.5, and 17% w/v PEG 10 000, and the G45V mutant crystallizes from 0.15 M ammonium acetate, 0.1 M Bis-Tris, pH 5.5, 3% w/v PEG 20 000, and 10 mM EDTA, 16°C, X-ray diffraction strutcure determination and analysis at 2.4 A, 3.3 A, and 2.7 A resolution, respectively	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant His-tagged wild-type and mutant enzymes, sitting drop vapour diffusion method, apo wild-type GlnRS crystallizes from 0.1 M calcium acetate, 0.1 M Tris, pH 6.0, 12.5% w/v PEG 3350, and 60 mM Gly-Gly-Gly, the Y57H mutant crystallizes from 0.1 M ammonium acetate, 0.1 M Bis-Tris, pH 5.5, and 17% w/v PEG 10 000, and the G45V mutant crystallizes from 0.15 M ammonium acetate, 0.1 M Bis-Tris, pH 5.5, 3% w/v PEG 20 000, and 10 mM EDTA, 16°C, X-ray diffraction strutcure determination and analysis at 2.4 A, 3.3 A, and 2.7 A resolution, respectively

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
G45V	naturally occuring mutation involved in development of brain disorder, modestly affects the conformation of the N-terminal domain and the stability of GlnRS, the mutant shows reduced activity compared to the wild-type. Gly45 is in the solvent-flexible loop between helices alpha4 and alpha5	Homo sapiens
H175A	the mutant shows reduced activity compared to the wild-type	Homo sapiens
additional information	generation of truncated mutants DELTAN1 (116-775) with reduced activity compared to the wild-type, and DELTANTD (183-775) with no activity	Homo sapiens
R403W	inactive mutant, the mutant is bound to GroEL when recombinantly expressed in Escherichi coli suggesting that it is misfolded	Homo sapiens
R515W	inactive mutant, the mutant is bound to GroEL when recombinantly expressed in Escherichi coli suggesting that it is misfolded	Homo sapiens
Y57H	naturally occuring mutation involved in development of brain disorder, modestly affects the conformation of the N-terminal domain and the stability of GlnRS, the mutant shows reduced activity compared to the wild-type. Tyr57 is in the middle of helix alpha5	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.00053	-	tRNAGln	recombinant mutant G45V, pH 7.2, 37°C	Homo sapiens
0.0006	-	tRNAGln	recombinant mutant H175A, pH 7.2, 37°C	Homo sapiens
0.00067	-	tRNAGln	recombinant wild-type enzyme, pH 7.2, 37°C	Homo sapiens
0.0017	-	tRNAGln	recombinant mutant Y57H, pH 7.2, 37°C	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cytosol	-	Homo sapiens	5829	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + L-glutamine + tRNAGln	Homo sapiens	-	AMP + diphosphate + L-glutaminyl-tRNAGln	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P47897	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta (DE3)pLysS by nickel affinity chromatography and gel filtration	Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining
brain	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + L-glutamine + tRNAGln	-	Homo sapiens	AMP + diphosphate + L-glutaminyl-tRNAGln	-	?
additional information	human GlnRS cannot aminoacylate bacterial tRNAGln from Escherichia coli, but recognition of tRNAGln by human and yeast GlnRSs may be conserved	Homo sapiens	?	-	?

Subunits

Subunits	Comment	Organism
monomer	the enzyme adopts a boomerang-like shaped structure built of 5 domains, domain organization of the intact enzyme and structure of the functionally important N-terminal domain, modeling of overall structure and domain organization of wild-type, full-length enzyme, structure-function analysis, overview	Homo sapiens

Synonyms

Synonyms	Comment	Organism
cytosolic glutaminyl-tRNA synthetase	-	Homo sapiens
GlnRS	-	Homo sapiens
Glutaminyl-tRNA synthetase	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
additional information	-	the melting curves of mutant enzymes G45V and Y57H comprise two peaks indicating that these mutants unfold along a more complex trajectory. The (un)folding events occur at 46.5°C and 52°C in G45V, and at 46°C and 51°C in Y57H	Homo sapiens
47.5	-	melting temperature of the recombinant wild-type enzyme	Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.002	-	tRNAGln	recombinant mutant Y57H, pH 7.2, 37°C	Homo sapiens
0.0033	-	tRNAGln	recombinant mutant H175A, pH 7.2, 37°C	Homo sapiens
0.0148	-	tRNAGln	recombinant mutant G45V, pH 7.2, 37°C	Homo sapiens
0.203	-	tRNAGln	recombinant wild-type enzyme, pH 7.2, 37°C	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.2	-	assay at	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Homo sapiens

General Information

General Information	Comment	Organism
evolution	human GlnRS is a monomeric class I aminoacyl-tRNA synthetase family member	Homo sapiens
malfunction	heterozygous mutations in GlnRS cause severe brain disorders. Pathological mutations mapping in the N-terminal domain alter the domain structure, and decrease catalytic activity and stability of GlnRS, whereas missense mutations in the catalytic domain induce misfolding of the enzyme. The reduced catalytic efficiency and a propensity of GlnRS mutants to misfold trigger the disease development	Homo sapiens
additional information	structure-function analysis, importance of the N-terminal domain for GlnRS function, overview	Homo sapiens
physiological function	cytosolic glutaminyl-tRNA synthetase (GlnRS) is the singular enzyme responsible for translation of glutamine codons	Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.118	-	tRNAGln	recombinant mutant Y57H, pH 7.2, 37°C	Homo sapiens
5.55	-	tRNAGln	recombinant mutant H175A, pH 7.2, 37°C	Homo sapiens
27.93	-	tRNAGln	recombinant mutant G45V, pH 7.2, 37°C	Homo sapiens
302.99	-	tRNAGln	recombinant wild-type enzyme, pH 7.2, 37°C	Homo sapiens