Literature summary for 6.1.1.2 extracted from

Shen, N.; Guo, L.; Yang, B.; Jin, Y.; Ding, J.
Structure of human tryptophanyl-tRNA synthetase in complex with tRNATrp reveals the molecular basis of tRNA recognition and specificity (2006), Nucleic Acids Res., 34, 3246-3258.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant TrpRSs in Escherichia coli	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme in complex with bovine tRNATrp, hanging drop vapor diffusion method, 4°C, 0.002 ml of protein solution containing 8 mg/ml of TrpRS with an equal amount of tRNA, 10 mM ATP, 1 mM Trp, 20 mM K2HPO4, pH 6.8, 10 mM MgCl2, 5 mM 2-mercaptoethanol and 0.5 mM PMSF, mixed with 0.002 ml reservoir solution containing 2 M (NH4)2SO4 and 50 mM HEPES, pH 7.0, 1-2 months, X-ray diffraction structure determination and analysis at 3.0 A resolution, modeling	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant enzyme in complex with bovine tRNATrp, hanging drop vapor diffusion method, 4°C, 0.002 ml of protein solution containing 8 mg/ml of TrpRS with an equal amount of tRNA, 10 mM ATP, 1 mM Trp, 20 mM K2HPO4, pH 6.8, 10 mM MgCl2, 5 mM 2-mercaptoethanol and 0.5 mM PMSF, mixed with 0.002 ml reservoir solution containing 2 M (NH4)2SO4 and 50 mM HEPES, pH 7.0, 1-2 months, X-ray diffraction structure determination and analysis at 3.0 A resolution, modeling

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
D99A	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
D99E	site-directed mutagenesis, the mutant shows slightly reduced activity and kcat compared to the wild-type enzyme	Homo sapiens
D99K	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
D99V	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
K102A	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
K102D	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
K102I	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
K102R	site-directed mutagenesis, the mutant shows slightly reduced activity and reduced kcat compared to the wild-type enzyme	Homo sapiens
K431A	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
K431D	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
K431I	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
K431R	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
Q194A	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
Q194L	site-directed mutagenesis, the mutant is inactive	Homo sapiens
R106A	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
R106D	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
R106I	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
R106K	site-directed mutagenesis, the mutant shows slightly reduced activity and reduced kcat compared to the wild-type enzyme	Homo sapiens
Y159A	site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme	Homo sapiens
Y159F	site-directed mutagenesis, the mutant shows slightly reduced activity and reduce kcat compared to the wild-type enzyme	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	kinetics of diverse recombinant mutant enzymes, overview	Homo sapiens
0.00121	-	tRNATrp	pH 7.5, 30°C, recombinant wild-type enzyme	Homo sapiens

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + L-tryptophan + tRNATrp	Homo sapiens	-	AMP + diphosphate + L-tryptophanyl-tRNATrp	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P23381	-	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	recombinant wild-type and mutant enzymes	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + L-tryptophan + tRNATrp	-	Homo sapiens	AMP + diphosphate + L-tryptophanyl-tRNATrp	-	?
ATP + L-tryptophan + tRNATrp	tRNA substrate from Bos taurus, a two step reaction: the amino acid is first activated by ATP to form an aminoacyl-AMP, which is then transferred to the 3' end of the cognate tRNA to form an aminoacyl-tRNA, the discriminator base A73 of the tRNA is specifically recognized by an alpha-helix of the unique N-terminal domain and the anticodon loop by an alpha-helix insertion of the C-terminal domain, the N-terminal domain is involved in Trp activation, but is not essential for tRNA binding and acylation, tryptophan, anticodon, and acceptor arm recognition mechanisms, overview	Homo sapiens	AMP + diphosphate + L-tryptophanyl-tRNATrp	-	?

Synonyms

Synonyms	Comment	Organism
TrpRS	-	Homo sapiens
Tryptophanyl-tRNA synthetase	-	Homo sapiens

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
1.33	-	tRNATrp	pH 7.5, 30°C, recombinant wild-type enzyme	Homo sapiens

pH Optimum

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

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Homo sapiens