UniProt functional annotation for P0AGJ9



	UniProt functional annotation for P0AGJ9

UniProt code: P0AGJ9.

Organism: Escherichia coli (strain K12).

Taxonomy: Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales; Enterobacteriaceae; Escherichia.

Function: Catalyzes the attachment of L-tyrosine to tRNA(Tyr) in a two- step reaction: tyrosine is first activated by ATP to form Tyr-AMP and then transferred to the acceptor end of tRNA(Tyr) (PubMed:4292198, PubMed:4579631). Can also mischarge tRNA(Tyr) with D-tyrosine, leading to the formation of D-tyrosyl-tRNA(Tyr), which can be hydrolyzed by the D-aminoacyl-tRNA deacylase (PubMed:4292198). In vitro, can also use the non-natural amino acid azatyrosine (PubMed:11006270). {ECO:0000269|PubMed:11006270, ECO:0000269|PubMed:4292198, ECO:0000269|PubMed:4579631}.

Catalytic activity: Reaction=ATP + L-tyrosine + tRNA(Tyr) = AMP + diphosphate + H(+) + L- tyrosyl-tRNA(Tyr); Xref=Rhea:RHEA:10220, Rhea:RHEA-COMP:9706, Rhea:RHEA-COMP:9707, ChEBI:CHEBI:15378, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:58315, ChEBI:CHEBI:78442, ChEBI:CHEBI:78536, ChEBI:CHEBI:456215; EC=6.1.1.1; Evidence={ECO:0000255|HAMAP-Rule:MF_02006, ECO:0000269|PubMed:10572925, ECO:0000269|PubMed:11006270, ECO:0000269|PubMed:4292198, ECO:0000269|PubMed:4579631};

Catalytic activity: Reaction=ATP + D-tyrosine + tRNA(Tyr) = AMP + D-tyrosyl-tRNA(Tyr) + diphosphate + H(+); Xref=Rhea:RHEA:57448, Rhea:RHEA-COMP:9707, Rhea:RHEA-COMP:9872, ChEBI:CHEBI:15378, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:58570, ChEBI:CHEBI:78442, ChEBI:CHEBI:78723, ChEBI:CHEBI:456215; Evidence={ECO:0000269|PubMed:4292198};

Activity regulation: Magnesium is essential for activity (PubMed:4579631). Inhibited by chloride and sulfate in the presence of 1 mM free Mg(2+). When the Mg(2+) concentration increases to 10 mM there is almost no chloride inhibition any more. Inhibited by diphosphate and AMP. Chloride strengthens the diphosphate inhibition and weakens the AMP inhibition. Chloride weakens the binding of Mg(2+) to the RNA and thereby the interaction between the enzyme and the RNA (PubMed:10572925). Acetylation at certain lysine residues could significantly impair activity (PubMed:28741290). D-tyrosine is a competitive inhibitor of L-tyrosine for the formation of tyrosyl- tRNA(Tyr) (PubMed:4292198). {ECO:0000269|PubMed:10572925, ECO:0000269|PubMed:28741290, ECO:0000269|PubMed:4292198, ECO:0000269|PubMed:4579631}.

Biophysicochemical properties: Kinetic parameters: KM=0.027 mM for L-tyrosine {ECO:0000269|PubMed:4292198}; KM=0.0033 mM for L-tyrosine {ECO:0000269|PubMed:11006270}; KM=0.015 mM for D-tyrosine {ECO:0000269|PubMed:4292198}; KM=0.0177 mM for azatyrosine {ECO:0000269|PubMed:11006270}; KM=22 nM for tRNA(Tyr) (in the absence of KCl) {ECO:0000269|PubMed:10572925}; KM=37 nM for tRNA(Tyr) (in the presence of 50 mM KCl) {ECO:0000269|PubMed:10572925}; KM=93 nM for tRNA(Tyr) (in the presence of 100 mM KCl) {ECO:0000269|PubMed:10572925}; KM=240 nM for tRNA(Tyr) (in the presence of 150 mM KCl) {ECO:0000269|PubMed:10572925}; Vmax=2.6 umol/min/mg enzyme for L-tyrosine {ECO:0000269|PubMed:4292198}; Vmax=0.11 umol/min/mg enzyme for D-tyrosine {ECO:0000269|PubMed:4292198}; Note=kcat is 0.74 sec(-1) with L-tyrosine as substrate. kcat is 0.11 sec(-1) with azatyrosine as substrate. {ECO:0000269|PubMed:11006270};

Subunit: Homodimer (PubMed:4579631, PubMed:6754952, PubMed:15663931, PubMed:15671170). Binds one molecule of tRNA(Tyr) per homodimer (PubMed:6754952). {ECO:0000269|PubMed:15663931, ECO:0000269|PubMed:15671170, ECO:0000269|PubMed:4579631, ECO:0000269|PubMed:6754952}.

Subcellular location: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_02006, ECO:0000305}.

Ptm: Acetylated at Lys-144 (PubMed:18723842, PubMed:28741290). Acetylation at Lys-144 leads to slightly decreased activity (PubMed:28741290). In vitro, in the presence of acetyl-phosphate, can also be acetylated at Lys-67, Lys-85, Lys-90, Lys-230, Lys-235, Lys- 238, Lys-321 and Lys-377. Acetylation at Lys-85, Lys-235 or Lys-238 causes dramatic decrease in activity (PubMed:28741290). {ECO:0000269|PubMed:18723842, ECO:0000269|PubMed:28741290}.

Biotechnology: Utilization for in vivo protein biosynthesis of mutants that charge azatyrosine efficiently to tRNA may lead to efficient production of azatyrosine-containing alloproteins, which have immense potential in biotechnology and medicine. {ECO:0000269|PubMed:11006270}.

Similarity: Belongs to the class-I aminoacyl-tRNA synthetase family. TyrS type 1 subfamily. {ECO:0000255|HAMAP-Rule:MF_02006, ECO:0000305}.

Annotations taken from UniProtKB at the EBI.


Organism:		Escherichia coli (strain K12).
Taxonomy:		Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales; Enterobacteriaceae; Escherichia.



Function:		Catalyzes the attachment of L-tyrosine to tRNA(Tyr) in a two- step reaction: tyrosine is first activated by ATP to form Tyr-AMP and then transferred to the acceptor end of tRNA(Tyr) (PubMed:4292198, PubMed:4579631). Can also mischarge tRNA(Tyr) with D-tyrosine, leading to the formation of D-tyrosyl-tRNA(Tyr), which can be hydrolyzed by the D-aminoacyl-tRNA deacylase (PubMed:4292198). In vitro, can also use the non-natural amino acid azatyrosine (PubMed:11006270). {ECO:0000269\|PubMed:11006270, ECO:0000269\|PubMed:4292198, ECO:0000269\|PubMed:4579631}.


Catalytic activity:		Reaction=ATP + L-tyrosine + tRNA(Tyr) = AMP + diphosphate + H(+) + L- tyrosyl-tRNA(Tyr); Xref=Rhea:RHEA:10220, Rhea:RHEA-COMP:9706, Rhea:RHEA-COMP:9707, ChEBI:CHEBI:15378, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:58315, ChEBI:CHEBI:78442, ChEBI:CHEBI:78536, ChEBI:CHEBI:456215; EC=6.1.1.1; Evidence={ECO:0000255\|HAMAP-Rule:MF_02006, ECO:0000269\|PubMed:10572925, ECO:0000269\|PubMed:11006270, ECO:0000269\|PubMed:4292198, ECO:0000269\|PubMed:4579631};
Catalytic activity:		Reaction=ATP + D-tyrosine + tRNA(Tyr) = AMP + D-tyrosyl-tRNA(Tyr) + diphosphate + H(+); Xref=Rhea:RHEA:57448, Rhea:RHEA-COMP:9707, Rhea:RHEA-COMP:9872, ChEBI:CHEBI:15378, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:58570, ChEBI:CHEBI:78442, ChEBI:CHEBI:78723, ChEBI:CHEBI:456215; Evidence={ECO:0000269\|PubMed:4292198};
Activity regulation:		Magnesium is essential for activity (PubMed:4579631). Inhibited by chloride and sulfate in the presence of 1 mM free Mg(2+). When the Mg(2+) concentration increases to 10 mM there is almost no chloride inhibition any more. Inhibited by diphosphate and AMP. Chloride strengthens the diphosphate inhibition and weakens the AMP inhibition. Chloride weakens the binding of Mg(2+) to the RNA and thereby the interaction between the enzyme and the RNA (PubMed:10572925). Acetylation at certain lysine residues could significantly impair activity (PubMed:28741290). D-tyrosine is a competitive inhibitor of L-tyrosine for the formation of tyrosyl- tRNA(Tyr) (PubMed:4292198). {ECO:0000269\|PubMed:10572925, ECO:0000269\|PubMed:28741290, ECO:0000269\|PubMed:4292198, ECO:0000269\|PubMed:4579631}.
Biophysicochemical properties:		Kinetic parameters: KM=0.027 mM for L-tyrosine {ECO:0000269\|PubMed:4292198}; KM=0.0033 mM for L-tyrosine {ECO:0000269\|PubMed:11006270}; KM=0.015 mM for D-tyrosine {ECO:0000269\|PubMed:4292198}; KM=0.0177 mM for azatyrosine {ECO:0000269\|PubMed:11006270}; KM=22 nM for tRNA(Tyr) (in the absence of KCl) {ECO:0000269\|PubMed:10572925}; KM=37 nM for tRNA(Tyr) (in the presence of 50 mM KCl) {ECO:0000269\|PubMed:10572925}; KM=93 nM for tRNA(Tyr) (in the presence of 100 mM KCl) {ECO:0000269\|PubMed:10572925}; KM=240 nM for tRNA(Tyr) (in the presence of 150 mM KCl) {ECO:0000269\|PubMed:10572925}; Vmax=2.6 umol/min/mg enzyme for L-tyrosine {ECO:0000269\|PubMed:4292198}; Vmax=0.11 umol/min/mg enzyme for D-tyrosine {ECO:0000269\|PubMed:4292198}; Note=kcat is 0.74 sec(-1) with L-tyrosine as substrate. kcat is 0.11 sec(-1) with azatyrosine as substrate. {ECO:0000269\|PubMed:11006270};
Subunit:		Homodimer (PubMed:4579631, PubMed:6754952, PubMed:15663931, PubMed:15671170). Binds one molecule of tRNA(Tyr) per homodimer (PubMed:6754952). {ECO:0000269\|PubMed:15663931, ECO:0000269\|PubMed:15671170, ECO:0000269\|PubMed:4579631, ECO:0000269\|PubMed:6754952}.
Subcellular location:		Cytoplasm {ECO:0000255\|HAMAP-Rule:MF_02006, ECO:0000305}.
Ptm:		Acetylated at Lys-144 (PubMed:18723842, PubMed:28741290). Acetylation at Lys-144 leads to slightly decreased activity (PubMed:28741290). In vitro, in the presence of acetyl-phosphate, can also be acetylated at Lys-67, Lys-85, Lys-90, Lys-230, Lys-235, Lys- 238, Lys-321 and Lys-377. Acetylation at Lys-85, Lys-235 or Lys-238 causes dramatic decrease in activity (PubMed:28741290). {ECO:0000269\|PubMed:18723842, ECO:0000269\|PubMed:28741290}.
Biotechnology:		Utilization for in vivo protein biosynthesis of mutants that charge azatyrosine efficiently to tRNA may lead to efficient production of azatyrosine-containing alloproteins, which have immense potential in biotechnology and medicine. {ECO:0000269\|PubMed:11006270}.
Similarity:		Belongs to the class-I aminoacyl-tRNA synthetase family. TyrS type 1 subfamily. {ECO:0000255\|HAMAP-Rule:MF_02006, ECO:0000305}.