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Literature summary for 6.1.1.1 extracted from

  • Hamano-Takaku, F.; Iwama, T.; Saito-Yano, S.; Takaku, K.; Monden, Y.; Kitabatake, M.; Soll, D.; Nishimura, S.
    A mutant Escherichia coli tyrosyl-tRNA synthetase utilizes the unnatural amino acid azatyrosine more efficiently than tyrosine (2000), J. Biol. Chem., 275, 40324-40328.
    View publication on PubMed
Search for more literature for 6.1.1.1

Cloned(Commentary)

Cloned (Comment) Organism
gene tyrS, expression of wild-type and mutant enzymes as His-tagged proteins in strain JM109 Escherichia coli

Protein Variants

Protein Variants Comment Organism
F130S construction of a plasmid library of randomly mutated gene tyrS by PCR, isolation of a mutant R-6-A-7 which incorporates L-beta-(5-hydroxy-2-pyridyl)-alanine in transformed Escherichia coli cells in vivo, increased temperature instability Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0033
-
 L-tyrosine wild-type enzyme, pH 7.5, 30°C Escherichia coli
0.018
-
 L-beta-(5-hydroxy-2-pyridyl)-alanine wild-type enzyme, pH 7.5, 30°C Escherichia coli
0.038
-
 L-beta-(5-hydroxy-2-pyridyl)-alanine mutant F130S, pH 7.5, 30°C Escherichia coli
0.066
-
 L-tyrosine mutant F130S, pH 7.5, 30°C Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-beta-(5-hydroxy-2-pyridyl)-alanine + tRNATyr Escherichia coli L-beta-(5-hydroxy-2-pyridyl)-alanine i.e. azatyrosine, mutant F130S shows 17fold higher activity in vivo than the wild-type enzyme AMP + L-beta-(5-hydroxy-2-pyridyl)-alanine-tRNATyr + diphosphate
-
 ?
ATP + L-tyrosine + tRNATyr Escherichia coli
-
 AMP + L-Tyr-tRNATyr + diphosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant HIs-tagged wild-type and mutant F130S from strain JM109 Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-beta-(5-hydroxy-2-pyridyl)-alanine + tRNATyr L-beta-(5-hydroxy-2-pyridyl)-alanine i.e. azatyrosine, mutant F130S, and wild-type enzyme the latter showing low activity Escherichia coli AMP + L-beta-(5-hydroxy-2-pyridyl)-alanine-tRNATyr + diphosphate
-
 ?
ATP + L-beta-(5-hydroxy-2-pyridyl)-alanine + tRNATyr L-beta-(5-hydroxy-2-pyridyl)-alanine i.e. azatyrosine, mutant F130S shows 17fold higher activity in vivo than the wild-type enzyme Escherichia coli AMP + L-beta-(5-hydroxy-2-pyridyl)-alanine-tRNATyr + diphosphate
-
 ?
ATP + L-tyrosine + tRNATyr
-
 Escherichia coli AMP + L-Tyr-tRNATyr + diphosphate
-
 ?

Subunits

Subunits Comment Organism
More three-dimensional modeling of the Escherichia coli enzyme constructed on the basis of the X-ray crystal structure of Bacillus stearothermophilus enzyme complexed with tyrosinyl adenylate, PDB code 3TS1 Escherichia coli

Synonyms

Synonyms Comment Organism
Tyrosine tRNA synthetase
-
 Escherichia coli
TyrRS
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
 10 min, wild-type enzyme is not affected, while the mutant F130S is almost completely inactivated Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.042
-
 L-beta-(5-hydroxy-2-pyridyl)-alanine mutant F130S, pH 7.5, 30°C Escherichia coli
0.11
-
 L-beta-(5-hydroxy-2-pyridyl)-alanine wild-type enzyme, pH 7.5, 30°C Escherichia coli
0.74
-
 L-tyrosine wild-type enzyme, pH 7.5, 30°C Escherichia coli
1.4
-
 L-tyrosine mutant F130S, pH 7.5, 30°C Escherichia coli
6.08
-
 L-tyrosine wild-type enzyme, pH 7.5, 30°C Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Escherichia coli