Literature summary extracted from
Caillet, J.; Graffe, M.; Eyermann, F.; Romby, P.; Springer, M.
Mutations in residues involved in zinc binding in the catalytic site of Escherichia coli threonyl-tRNA synthetase confer a dominant lethal phenotype (2007), J. Bacteriol., 189, 6839-6848.
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
6.1.1.3 |
overexpression of His-tagged mutant enzymes |
Escherichia coli K-12 |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
6.1.1.3 |
H385A |
site-directed mutagenesis, the mutant shows altered substrate specificity, overview |
Escherichia coli K-12 |
6.1.1.3 |
H385N |
site-directed mutagenesis, the mutant shows altered substrate specificity, overview |
Escherichia coli K-12 |
6.1.1.3 |
H385Y |
site-directed mutagenesis, the mutant shows altered substrate specificity, overview |
Escherichia coli K-12 |
6.1.1.3 |
additional information |
mutations of any of the three amino acids forming the zinc-binding site inactivate the enzyme and have a dominant negative effect on growth if the corresponding genes are placed on a multicopy plasmid, not due to the formation of inactive heterodimers, the titration of tRNAThr by an inactive enzyme, or its misaminoacylation but is, rather, due to the regulatory function of threonyl-tRNA synthetase, overview, the mutations confer a dominant lethal phenotype, overproduction of the inactive enzyme represses the expression of the wild-type chromosomal copy of the gene to an extent incompatible with bacterial growth, phenotypes, overview |
Escherichia coli K-12 |
6.1.1.3 |
R583H |
site-directed mutagenesis, the mutant shows altered substrate specificity, overview |
Escherichia coli K-12 |
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
6.1.1.3 |
Mg2+ |
- |
Escherichia coli K-12 |
|
6.1.1.3 |
Zn2+ |
zinc binding in the catalytic site, the active site zinc atom is essential for the recognition of threonine, three amino acids forming the zinc-binding site, overview |
Escherichia coli K-12 |
|
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
6.1.1.3 |
ATP + L-threonine + tRNAThr |
Escherichia coli K-12 |
the enzyme acts as both an enzyme and a regulator of gene expression, it aminoacylates tRNAThr isoacceptors and binds to its own mRNA, inhibiting its translation, overview |
AMP + diphosphate + L-threonyl-tRNAThr |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
6.1.1.3 |
Escherichia coli K-12 |
- |
several strains, overview, gene thrS |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
6.1.1.3 |
recombinant His-tagged mutant H385A |
Escherichia coli K-12 |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
6.1.1.3 |
ATP + L-threonine + tRNAThr |
the enzyme acts as both an enzyme and a regulator of gene expression, it aminoacylates tRNAThr isoacceptors and binds to its own mRNA, inhibiting its translation, overview |
Escherichia coli K-12 |
AMP + diphosphate + L-threonyl-tRNAThr |
- |
? |
|
6.1.1.3 |
ATP + L-threonine + tRNAThr |
the zinc atom in the active site is essential for the recognition of threonine |
Escherichia coli K-12 |
AMP + diphosphate + L-threonyl-tRNAThr |
- |
? |
|
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
6.1.1.3 |
Threonyl-tRNA synthetase |
- |
Escherichia coli K-12 |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
6.1.1.3 |
7.5 |
- |
assay at |
Escherichia coli K-12 |
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
6.1.1.3 |
ATP |
- |
Escherichia coli K-12 |
|