Cloned (Comment) | Organism |
---|---|
expression of an N-terminally His6-tagged MetRS monomer in Escherichia coli strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D369A | site-directed mutagenesis in the MetRS SCF, the mutant shows reduced transfer RNA aminoacylation and 125fold loss in tRNAMet aminoacylation efficiency compared to the wild-type enzyme | Escherichia coli |
D369K/K295D | site-directed mutagenesis in the MetRS SCF, the mutant shows reduced transfer RNA aminoacylation compared to the wild-type enzyme | Escherichia coli |
D369N | site-directed mutagenesis in the MetRS SCF, the mutant shows reduced transfer RNA aminoacylation and 60fold loss in tRNAMet aminoacylation efficiency compared to the wild-type enzyme | Escherichia coli |
K295A | site-directed mutagenesis in the MetRS SCF, the mutant shows reduced transfer RNA aminoacylation compared to the wild-type enzyme | Escherichia coli |
K295V | site-directed mutagenesis in the MetRS SCF, the mutant shows reduced transfer RNA aminoacylation compared to the wild-type enzyme | Escherichia coli |
additional information | replacement of amino acids of the MetRS SCF with portions of the structurally similar glutaminyl-tRNA synthetase, EC6.1.1.18, motif or with alanine residues. Chimeric variants retain significant tRNA methionylation activity, indicating that structural integrity of the helix-turn-strand-helix motif contributes more to RNA aminoacylation than does amino acid identity. In contrast, chimeras are significantly reduced in methionyl adenylate synthesis, suggesting a role for the SCF in formation of a structured active site domain | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic parameters for tRNAMet aminoacylation and for MetRS-catalyzed diphosphate exchange of wild-type enzyme and mutants, overview | Escherichia coli | |
0.0039 | - |
tRNAMet | pH 7.5, 25°C, wild-type enzyme | Escherichia coli | |
0.006 | - |
tRNAMet | pH 7.5, 25°C, mutant D369K/K295D | Escherichia coli | |
0.0102 | - |
tRNAMet | pH 7.5, 25°C, mutant KA295V | Escherichia coli | |
0.0177 | - |
tRNAMet | pH 7.5, 25°C, mutant K295V | Escherichia coli | |
0.018 | - |
tRNAMet | pH 7.5, 25°C, mutant D369A | Escherichia coli | |
0.026 | - |
tRNAMet | pH 7.5, 25°C, mutant D369N | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-methionine + tRNAMet | Escherichia coli | - |
AMP + diphosphate + L-methionyl-tRNAMet | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His6-tagged MetRS monomer from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-methionine + tRNAMet | - |
Escherichia coli | AMP + diphosphate + L-methionyl-tRNAMet | - |
? |
Synonyms | Comment | Organism |
---|---|---|
MetRS | - |
Escherichia coli |
More | the enzyme is a class I aminoacyl-tRNA synthetase | Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.037 | - |
tRNAMet | pH 7.5, 25°C, mutant D369A | Escherichia coli | |
0.042 | - |
tRNAMet | pH 7.5, 25°C, mutant KA295V | Escherichia coli | |
0.05 | - |
tRNAMet | pH 7.5, 25°C, mutant D369K/K295D | Escherichia coli | |
0.11 | - |
tRNAMet | pH 7.5, 25°C, mutant D369N | Escherichia coli | |
0.3 | - |
tRNAMet | pH 7.5, 25°C, mutant K295V | Escherichia coli | |
1 | - |
tRNAMet | pH 7.5, 25°C, wild-type enzyme | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli |
General Information | Comment | Organism |
---|---|---|
additional information | the catalytic domains of class I aminoacyl-tRNA synthetases are built around a conserved Rossmann nucleotide binding fold, with additional polypeptide domains responsible for tRNA binding or hydrolytic editing of misacylated substrates, structural comparisons of class Ia and Ib enzymes, overview. Structural integrity of the helix-turn-strand-helix motif contributes more to tRNA aminoacylation than does amino acid identity | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | - |
tRNAMet | pH 7.5, 25°C, mutant D369A | Escherichia coli | |
4 | - |
tRNAMet | pH 7.5, 25°C, mutant D369N | Escherichia coli | |
4 | - |
tRNAMet | pH 7.5, 25°C, mutant KA295V | Escherichia coli | |
8 | - |
tRNAMet | pH 7.5, 25°C, mutant D369K/K295D | Escherichia coli | |
17 | - |
tRNAMet | pH 7.5, 25°C, mutant K295V | Escherichia coli | |
250 | - |
tRNAMet | pH 7.5, 25°C, wild-type enzyme | Escherichia coli |