Cloned (Comment) | Organism |
---|---|
gene thrS, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Escherichia coli |
gene thrS, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Mesomycoplasma mobile |
gene thrS, sequence comparisons, recombinant expression of wild-type enzyme in Escherichia coli strain BL21(DE3) | Mycoplasma capricolum |
Protein Variants | Comment | Organism |
---|---|---|
D46E | site-directed mutagenesis, the mutant has a modest reduction in its aminoacylation activity compared to wild-type, and its post-transfer editing activity is abolished | Escherichia coli |
D46E/H186G | site-directed mutagenesis, the mutant EcThrRS strongly supports growth of a yeast thrS deletion strain (ScDELTAthrS) | Escherichia coli |
D46E/Y173F | site-directed mutagenesis, the mutant EcThrRS supports growth of a yeast thrS deletion strain (ScDELTAthrS) | Escherichia coli |
D46E/Y173H | site-directed mutagenesis, the mutant does not support growth of a yeast thrS deletion strain (ScDELTAthrS) | Escherichia coli |
D46E/Y173K | site-directed mutagenesis, the mutant does not support growth of a yeast thrS deletion strain (ScDELTAthrS) | Escherichia coli |
D46E/Y173R | site-directed mutagenesis, the mutant does not support growth of a yeast thrS deletion strain (ScDELTAthrS) | Escherichia coli |
D46E/Y173S | site-directed mutagenesis, the mutant does not support growth of a yeast thrS deletion strain (ScDELTAthrS) | Escherichia coli |
D46R | site-directed mutagenesis, the mutant has a modest reduction in its aminoacylation activity compared to wild-type, and its post-transfer editing activity is abolished | Escherichia coli |
H186G | site-directed mutagenesis, the mutant EcThrRS strongly supports growth of a yeast thrS deletion strain (ScDELTAthrS) | Escherichia coli |
H9A/H13A | site-directed mutagenesis, the post-transfer editing of MmThrRS mutant H9A/H13A is reduced compared with that of wild-type MmThrRS, the in vivo mutation of more crucial residues is required to abolish the post-transfer editing | Mesomycoplasma mobile |
H9A/H13A/K86A/D117A/C119A/H123A | site-directed mutagenesis, the post-transfer editing of mutant MmThrRS-N2M is completely lost | Mesomycoplasma mobile |
K136A | site-directed mutagenesis, the mutant EcThrRS supports growth of a yeast thrS deletion strain (ScDELTAthrS) | Escherichia coli |
K136E | site-directed mutagenesis, the mutant does not support growth of a yeast thrS deletion strain (ScDELTAthrS) | Escherichia coli |
K136R | site-directed mutagenesis, the mutant EcThrRS supports growth of a yeast thrS deletion strain (ScDELTAthrS) | Escherichia coli |
additional information | the mutant EcThrRS-DELTAN1 lacking domain N1 shows no post-transfer editing activity | Escherichia coli |
Y173D | site-directed mutagenesis, the mutant has a modest reduction in its aminoacylation activity compared to wild-type, and its post-transfer editing activity is abolished | Escherichia coli |
Y173R | site-directed mutagenesis, the mutant has a modest reduction in its aminoacylation activity compared to wild-type, and its post-transfer editing activity is abolished | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of the enzyme for cognate Thr and noncognate Ser are determined with an ATP-phosphate exchange reaction | Mycoplasma capricolum | |
additional information | - |
additional information | kinetics of the enzyme for cognate Thr and noncognate Ser are determined with an ATP-phosphate exchange reaction | Mesomycoplasma mobile | |
3.39 | - |
L-threonine | pH 7.5, 30°C | Mycoplasma capricolum | |
4.4 | - |
L-threonine | pH 7.5, 30°C | Mesomycoplasma mobile | |
582.37 | - |
L-serine | pH 7.5, 30°C | Mycoplasma capricolum | |
939.72 | - |
L-serine | pH 7.5, 30°C | Mesomycoplasma mobile |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli | |
Mg2+ | required | Mesomycoplasma mobile |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-serine + tRNAThr | Mycoplasma capricolum | reaction of EC 6.1.1.11, mischarging of tRNAThr | AMP + diphosphate + L-seryl-tRNAThr | - |
? | |
ATP + L-serine + tRNAThr | Mesomycoplasma mobile | reaction of EC 6.1.1.11, mischarging of tRNAThr, low activity | AMP + diphosphate + L-seryl-tRNAThr | - |
? | |
ATP + L-serine + tRNAThr | Mesomycoplasma mobile ATCC 43663 | reaction of EC 6.1.1.11, mischarging of tRNAThr, low activity | AMP + diphosphate + L-seryl-tRNAThr | - |
? | |
ATP + L-threonine + tRNAThr | Mycoplasma capricolum | - |
AMP + diphosphate + L-threonyl-tRNAThr | - |
? | |
ATP + L-threonine + tRNAThr | Escherichia coli | - |
AMP + diphosphate + L-threonyl-tRNAThr | - |
? | |
ATP + L-threonine + tRNAThr | Mesomycoplasma mobile | - |
AMP + diphosphate + L-threonyl-tRNAThr | - |
? | |
ATP + L-threonine + tRNAThr | Mesomycoplasma mobile ATCC 43663 | - |
AMP + diphosphate + L-threonyl-tRNAThr | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A8M3 | - |
- |
Mesomycoplasma mobile | Q6KH76 | - |
- |
Mesomycoplasma mobile ATCC 43663 | Q6KH76 | - |
- |
Mycoplasma capricolum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) | Escherichia coli |
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) | Mesomycoplasma mobile |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-serine + tRNAThr | reaction of EC 6.1.1.11, mischarging of tRNAThr | Mycoplasma capricolum | AMP + diphosphate + L-seryl-tRNAThr | - |
? | |
ATP + L-serine + tRNAThr | reaction of EC 6.1.1.11, mischarging of tRNAThr, low activity | Mesomycoplasma mobile | AMP + diphosphate + L-seryl-tRNAThr | - |
? | |
ATP + L-serine + tRNAThr | reaction of EC 6.1.1.11, mischarging of tRNAThr, low activity | Mesomycoplasma mobile ATCC 43663 | AMP + diphosphate + L-seryl-tRNAThr | - |
? | |
ATP + L-threonine + tRNAThr | - |
Mycoplasma capricolum | AMP + diphosphate + L-threonyl-tRNAThr | - |
? | |
ATP + L-threonine + tRNAThr | - |
Escherichia coli | AMP + diphosphate + L-threonyl-tRNAThr | - |
? | |
ATP + L-threonine + tRNAThr | - |
Mesomycoplasma mobile | AMP + diphosphate + L-threonyl-tRNAThr | - |
? | |
ATP + L-threonine + tRNAThr | - |
Mesomycoplasma mobile ATCC 43663 | AMP + diphosphate + L-threonyl-tRNAThr | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | enzyme domain structure, threonyl-tRNA synthetase (ThrRS) contains multiple domains, including an N2 editing domain, overview | Mycoplasma capricolum |
More | enzyme domain structure, threonyl-tRNA synthetase (ThrRS) contains multiple domains, including an N2 editing domain, overview | Escherichia coli |
More | enzyme domain structure, threonyl-tRNA synthetase (ThrRS) contains multiple domains, including an N2 editing domain, overview | Mesomycoplasma mobile |
Synonyms | Comment | Organism |
---|---|---|
EcThrRS | - |
Escherichia coli |
McThrRS | - |
Mycoplasma capricolum |
MmThrRS | - |
Mesomycoplasma mobile |
Threonyl-tRNA synthetase | - |
Mycoplasma capricolum |
Threonyl-tRNA synthetase | - |
Escherichia coli |
Threonyl-tRNA synthetase | - |
Mesomycoplasma mobile |
ThrRS | - |
Mycoplasma capricolum |
ThrRS | - |
Escherichia coli |
ThrRS | - |
Mesomycoplasma mobile |
ThrS | - |
Mycoplasma capricolum |
ThrS | - |
Escherichia coli |
ThrS | - |
Mesomycoplasma mobile |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Mycoplasma capricolum |
30 | - |
assay at | Escherichia coli |
30 | - |
assay at | Mesomycoplasma mobile |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.55 | - |
L-serine | pH 7.5, 30°C | Mesomycoplasma mobile | |
1.15 | - |
L-serine | pH 7.5, 30°C | Mycoplasma capricolum | |
2.16 | - |
L-threonine | pH 7.5, 30°C | Mesomycoplasma mobile | |
6.31 | - |
L-threonine | pH 7.5, 30°C | Mycoplasma capricolum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Mycoplasma capricolum |
7.5 | - |
assay at | Escherichia coli |
7.5 | - |
assay at | Mesomycoplasma mobile |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli | |
ATP | - |
Mesomycoplasma mobile |
General Information | Comment | Organism |
---|---|---|
evolution | crucial importance of the only absolutely conserved residue within the N1 domain in regulating post-transfer editing activityand editing active sites by mediating an N1-N2 domain interaction in Escherichia coli ThrRS. Translational quality control of various ThrRSs and the role of the N1 domain in translational fidelity, overview | Escherichia coli |
evolution | ThrRSs from Mycoplasma species exhibit differences in their domain composition and editing active sites compared with the canonical ThrRSs. The Mycoplasma capricolum ThrRS, which harbors an N1 domain and a degenerate N2 domain, is editing-defective, and is thus not capable to support the growth of a yeast thrS deletion strain (ScDELTAthrS). Translational quality control of various ThrRSs and the role of the N1 domain in translational fidelity, overview. McThrRS has lost its post-transfer editing activity, probably because of its degenerate editing active site in the N2 domain. MmThrRS has negligible tRNA-dependent pretransfer editing capacities | Mycoplasma capricolum |
evolution | ThrRSs from Mycoplasma species exhibit differences in their domain composition and editing active sites compared with the canonical ThrRSs. The Mycoplasma mobile ThrRS, the first example of a ThrRS naturally lacking the N1 domain, displays efficient post-transfer editing activity. Mycoplasma mobile ThrRS is able to support the growth of a yeast thrS deletion strain (ScDELTAthrS). Translational quality control of various ThrRSs and the role of the N1 domain in translational fidelity, overview | Mesomycoplasma mobile |
additional information | structure analysis of EcThrRS, functional importance of the Asp46 in the N1 domain and the Tyr173 in the N2 domain | Escherichia coli |
physiological function | correct aminoacyl-tRNA generation is critical for the faithful transduction of genetic information, which is supported by the high levels of amino acid conservation in editing active sites of specific aaRSs across the three domains of life. Enzyme EcThrRS is an editing-capable enzyme, that can remove noncognate Ser. The N1 domain is essential for editing by EcThrRS. The enzyme shows strong tRNA-dependent editing, including the pre- and post-transfer editing of EcThrRS | Escherichia coli |
physiological function | correct aminoacyl-tRNA generation is critical for the faithful transduction of genetic information, which is supported by the high levels of amino acid conservation in editing active sites of specific aaRSs across the three domains of life. The enzyme misactivates noncognate Ser and therefore requires an editing function to ensure the correct Thr-tRNAThr formation. Enzyme McThrRS is not able to hydrolyze Ser-tRNAThr and remove noncognate Ser. The degeneration of the crucial editing active sites of McThrRS impairs its posttransfer editing, McThrRS has lost its post-transfer editing activity, probably because of its degenerate editing active site in the N2 domain. McThrRS is unable to complement the loss of Saccharomyces cerevisiae thrS in vivo, because of its lack of editing activity | Mycoplasma capricolum |
physiological function | correct aminoacyl-tRNA generation is critical for the faithful transduction of genetic information, which is supported by the high levels of amino acid conservation in editing active sites of specific aaRSs across the three domains of life. The enzyme misactivates noncognate Ser and therefore requires an editing function to ensure the correct Thr-tRNAThr formation. Enzyme MmThrRS is able to hydrolyze Ser-tRNAThr and remove noncognate Ser. MmThrRS, which has an intact N2 domain, has post-transfer editing activity, editing-crucial residues include Lys86, Asp117, Cys119, and His123. MmThrRS has negligible tRNA-dependent pretransfer editing capacities. MmThrRS can complement the loss of Saccharomyces cerevisiae thrS in vivo, because of its lack of editing activity. The absence of the N1 domain of MmThrRS increases in vivo activity and optimizes enzyme protein structure/stability | Mesomycoplasma mobile |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00059 | - |
L-serine | pH 7.5, 30°C | Mesomycoplasma mobile | |
0.00122 | - |
L-serine | pH 7.5, 30°C | Mycoplasma capricolum | |
0.49 | - |
L-threonine | pH 7.5, 30°C | Mesomycoplasma mobile | |
1.86 | - |
L-threonine | pH 7.5, 30°C | Mycoplasma capricolum |