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Literature summary extracted from

  • Senger, B.; Despons, L.; Walter, P.; Jakubowski, H.; Fasiolo, F.
    Yeast cytoplasmic and mitochondrial methionyl-tRNA synthetases: two structural frameworks for identical functions (2001), J. Mol. Biol., 311, 205-216.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.1.1.10 expression of cytoplasmic and mitochondrial isozymes as GST-fusion proteins in the yeast strain RS453, functional complementation of an inactive gene disruption yeast strain by the mitochondrial isozyme located in the cytoplasm Saccharomyces cerevisiae

Protein Variants

EC Number Protein Variants Comment Organism
6.1.1.10 A355C site-directed mutagenesis, 115% activity compared to the wild-type enzyme, in vivo complementation of a deficient yeast strain Saccharomyces cerevisiae
6.1.1.10 C337A/C340A site-directed mutagenesis, inactive Saccharomyces cerevisiae
6.1.1.10 C350A site-directed mutagenesis, 1.5% activity compared to the wild-type enzyme, no in vivo complementation of a deficient yeast strain Saccharomyces cerevisiae
6.1.1.10 C350A/C353A site-directed mutagenesis, inactive Saccharomyces cerevisiae
6.1.1.10 C353A site-directed mutagenesis, catalytically inactive, no in vivo complementation of a deficient yeast strain Saccharomyces cerevisiae
6.1.1.10 C367A site-directed mutagenesis, catalytically inactive, no in vivo complementation of a deficient yeast strain, mutant shows a second zinc-binding knuckle structure Saccharomyces cerevisiae
6.1.1.10 D348G site-directed mutagenesis, 4.7% activity compared to the wild-type enzyme, in vivo complementation of a deficient yeast strain Saccharomyces cerevisiae
6.1.1.10 D370A site-directed mutagenesis, 8.7% activity compared to the wild-type enzyme, in vivo complementation of a deficient yeast strain Saccharomyces cerevisiae
6.1.1.10 G347R site-directed mutagenesis, catalytically inactive, no in vivo complementation of a deficient yeast strain Saccharomyces cerevisiae
6.1.1.10 I363N site-directed mutagenesis, 84% activity compared to the wild-type enzyme, in vivo complementation of a deficient yeast strain Saccharomyces cerevisiae
6.1.1.10 additional information construction of an inactive strain by gene disruption Saccharomyces cerevisiae
6.1.1.10 P338I site-directed mutagenesis, 74% activity compared to the wild-type enzyme, in vivo complementation of a deficient yeast strain Saccharomyces cerevisiae

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.1.1.10 1,10-phenanthroline complete inhibition of the cytoplasmic isozyme at 1 mM Saccharomyces cerevisiae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.1.1.10 0.0004
-
 tRNAMet recombinant mitochondrial isozyme as GST-fusion protein, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 0.009
-
 tRNAMet recombinant cytoplasmic isozyme, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 0.015
-
 tRNAMet recombinant cytoplasmic isozyme as GST-fusion protein, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 1.2
-
 CoA methionylation reaction, cytoplasmic isozyme, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 1.8
-
 L-homocysteine editing reaction, mitochondrial isozyme, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 3.1
-
 L-homocysteine editing reaction, cytoplasmic isozyme, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 6.3
-
 CoA methionylation reaction, mitochondrial isozyme, pH 7.8, 25°C Saccharomyces cerevisiae

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
6.1.1.10 cytoplasm cytoplasmic isozyme has a zinc-finger motif and an insertion domain that divides the nucleotide-binding fold into 2 halves Saccharomyces cerevisiae 5737
-
6.1.1.10 mitochondrion mitochondrial isozyme contains no zinc-finger motif and no insertion domain in the nucleotide-binding fold Saccharomyces cerevisiae 5739
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.1.1.10 Zn2+ 2 binding sites, 0.14 mM tightly bound zinc is present in 1 mol of cytoplasmic isozyme, but not in the mitochondrial one, required for stability and activity of the cytoplasmic isozyme Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.1.1.10 ATP + L-homocysteine + tRNAMet Saccharomyces cerevisiae edition and aminoacylation by cytoplasmic and mitochondrial isozyme ?
-
 ?
6.1.1.10 ATP + L-methionine + tRNAMet Saccharomyces cerevisiae initiator and elongation tRNAMet, cytoplasmic and mitochondrial isozyme AMP + diphosphate + L-methionyl-tRNAMet
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.1.1.10 Saccharomyces cerevisiae
-
 2 cytoplasmic and mitochondrial isozymes
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.1.1.10 recombinant GST-fusion protein of the mitochondrial isozyme Saccharomyces cerevisiae

Reaction

EC Number Reaction Comment Organism Reaction ID
6.1.1.10 ATP + L-methionine + tRNAMet = AMP + diphosphate + L-methionyl-tRNAMet amino acid residues D138, R139, and F135 are important for activity Saccharomyces cerevisiae
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.1.1.10 ATP + L-homocysteine + tRNAMet edition and aminoacylation by cytoplasmic and mitochondrial isozyme Saccharomyces cerevisiae ?
-
 ?
6.1.1.10 ATP + L-methionine + tRNAMet initiator and elongation tRNAMet, cytoplasmic and mitochondrial isozyme Saccharomyces cerevisiae AMP + diphosphate + L-methionyl-tRNAMet
-
 ?
6.1.1.10 CoA + L-methionine
-
 Saccharomyces cerevisiae Met-S-CoA
-
 ?
6.1.1.10 additional information the isozymes also perform the ATP-diphosphate exchange reaction Saccharomyces cerevisiae ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.1.1.10 Methionine translase
-
 Saccharomyces cerevisiae
6.1.1.10 Methionine--tRNA ligase
-
 Saccharomyces cerevisiae
6.1.1.10 Methionyl tRNA synthetase
-
 Saccharomyces cerevisiae
6.1.1.10 Methionyl-transfer ribonucleate synthetase
-
 Saccharomyces cerevisiae
6.1.1.10 Methionyl-transfer ribonucleic acid synthetase
-
 Saccharomyces cerevisiae
6.1.1.10 Methionyl-transfer RNA synthetase
-
 Saccharomyces cerevisiae
6.1.1.10 MetRS
-
 Saccharomyces cerevisiae
6.1.1.10 Synthetase, methionyl-transfer ribonucleate
-
 Saccharomyces cerevisiae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.1.1.10 0.2
-
 tRNAMet recombinant mitochondrial isozyme as GST-fusion protein, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 0.5
-
 L-homocysteine editing reaction, mitochondrial isozyme, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 0.69
-
 L-homocysteine editing reaction, cytoplasmic isozyme, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 0.7
-
 CoA methionylation reaction, cytoplasmic isozyme, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 1
-
 tRNAMet recombinant cytoplasmic isozyme as GST-fusion protein, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 1.3
-
 tRNAMet recombinant cytoplasmic isozyme, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 1.6
-
 CoA methionylation reaction, mitochondrial isozyme, pH 7.8, 25°C Saccharomyces cerevisiae
6.1.1.10 6.08
-
 L-homocysteine editing reaction, cytoplasmic isozyme, pH 7.8, 25°C Saccharomyces cerevisiae

Cofactor

EC Number Cofactor Comment Organism Structure
6.1.1.10 ATP
-
 Saccharomyces cerevisiae