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Literature summary for 2.7.1.160 extracted from
- Substrate recognition by a yeast 2'-phosphotransferase involved in tRNA splicing and by its Escherichia coli homolog (2001), Biochemistry, 40, 14098-14105.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression in Escherichia coli BL21(DE3) | Escherichia coli |
| overexpression in Escherichia coli BL21(DE3) | Saccharomyces cerevisiae |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| ApGpApUpUpUpApC | oligonucleotide, which lacks secondary structure and has no terminal phosphate | Saccharomyces cerevisiae |  |
| [pre-tRNA] | intron-containing yeast [pre-tRNA], which has the same structure as mature yeast [tRNA], except in the region of the intron | Saccharomyces cerevisiae | |
| [tRNA] | mature Escherichia coli [tRNA], has canonical tRNA structue | Saccharomyces cerevisiae |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00000035 | - |
2'-phospho-[ligated tRNA] | - |
Saccharomyces cerevisiae | |
| 0.0000017 | - |
18-mer 2' phosphorylated RNA | pACAAGACUCUAA-(2-phosphate)AUCUUG, 30°C | Saccharomyces cerevisiae | |
| 0.000004 | - |
2'-phospho-[ligated tRNA] | - |
Escherichia coli | |
| 0.00002 | - |
8-mer 2' phosphorylated RNA | pGUAA-(2-phosphate)AUCU, 30°C | Saccharomyces cerevisiae | |
| 0.00015 | - |
18-mer 2' phosphorylated RNA | 37°C | Escherichia coli | |
| 0.00015 | - |
3-mer 2' phosphorylated RNA | pApA-(2-phosphate)pA, 30°C | Saccharomyces cerevisiae | |
| 0.00025 | - |
8-mer 2' phosphorylated RNA | 37°C | Escherichia coli | |
| 0.000325 | - |
3-mer 2' phosphorylated RNA | pUpU-(2-phosphate)pUs, 30°C | Saccharomyces cerevisiae | |
| 0.00048 | - |
3-mer 2' phosphorylated RNA | pApU-(2-phosphate)pA, 30°C | Saccharomyces cerevisiae | |
| 0.00065 | - |
3-mer 2' phosphorylated RNA | pApApA-(2-phosphate)p, 30°C | Saccharomyces cerevisiae | |
| 0.0007 | - |
3-mer 2' phosphorylated RNA | pApA-(2-phosphate)pA, 37°C | Escherichia coli | |
| 0.0013 | - |
3-mer 2' phosphorylated RNA | pAAA-(2-phosphate)p, 37°C | Escherichia coli | |
| 0.0018 | - |
3-mer 2' phosphorylated RNA | pUpU-(2-phosphate)pU, 37°C | Escherichia coli | |
| 0.003 | - |
3-mer 2' phosphorylated RNA | pApApA-(2-phosphate)pOCH3, 30°C | Saccharomyces cerevisiae | |
| 0.0038 | - |
3-mer 2' phosphorylated RNA | pAAA-(2-phosphate)pOCH3, 37°C | Escherichia coli | |
| 0.04 | - |
3-mer 2' phosphorylated RNA | pApApA-(2-phosphate), 30°C | Saccharomyces cerevisiae |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 30000 | - |
SDS-PAGE | Saccharomyces cerevisiae |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
- |
- |
| Saccharomyces cerevisiae | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| KptA-His6 10% purified by sonication, immobilized metal ion chromatography, centrifugation | Escherichia coli |
| Tpt1-His6 50% purified by sonication, immobilized metal ion chromatography, centrifugation | Saccharomyces cerevisiae |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
200000000 units/mg for recombinant Tpt1 produced in Escherichia coli and 140000000 units/mg for Tpt1 purified from yeast | Saccharomyces cerevisiae |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -20°C, Tris buffer, pH 7.5, 50% glycerol | Escherichia coli |
| -20°C, Tris buffer, pH 7.5, 50% glycerol | Saccharomyces cerevisiae |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 18-mer 2' phosphorylated RNA | - |
Escherichia coli | ? | - |
? | |
| 18-mer 2' phosphorylated RNA | pACAAGACUCUAA-(2-phosphate)AUCUUG | Saccharomyces cerevisiae | ? | - |
? | |
| 2'-phospho-[ligated tRNA] | - |
Escherichia coli | ? | - |
? | |
| 2'-phospho-[ligated tRNA] | - |
Saccharomyces cerevisiae | ? | - |
? | |
| 2'-phospho-[ligated tRNA] + NAD+ | - |
Escherichia coli | mature tRNA + ADP-ribose 1'',2''-phosphate + nicotinamide | - |
? | |
| 3-mer 2' phosphorylated RNA | pApA-(2-phosphate)pA, pApU-(2-phosphate)pA, pUpU-(2-phosphate)pU, pApApA-(2-phosphate),pApApA-(2-phosphate)p, pApApA-(2-phosphate)1OCH3 | Saccharomyces cerevisiae | ? | - |
? | |
| 3-mer 2' phosphorylated RNA | pApa-(2-phosphate)pA, pUpU-(2-phosphate)pU, pAAA-(2-phosphate)p and pAAA-(2-phosphate)pOCH3 | Escherichia coli | ? | - |
? | |
| 8-mer 2' phosphorylated RNA | - |
Escherichia coli | ? | - |
? | |
| 8-mer 2' phosphorylated RNA | pGUAA-(2-phosphate)AUCU | Saccharomyces cerevisiae | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| KptA | - |
Escherichia coli |
| Tpt1 | - |
Saccharomyces cerevisiae |
| yeast 2'-phosphotransferase | - |
Saccharomyces cerevisiae |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
8-mer 2' phosphorylated RNA | 37°C, vmax = 170 fmol/min/ng | Escherichia coli | |
| additional information | - |
18-mer 2' phosphorylated RNA | 37°C, vmax = 99 fmol/min/ng | Escherichia coli | |
| additional information | - |
3-mer 2' phosphorylated RNA | pAAA-(2-phosphate)p, 37°C, vmax = 1.4 fmol/min/ng | Escherichia coli | |
| additional information | - |
3-mer 2' phosphorylated RNA | pAAA-(2-phosphate)pOCH3, 37°C, vmax = 45 fmol/min/ng | Escherichia coli | |
| additional information | - |
3-mer 2' phosphorylated RNA | pApA-(2-phosphate)pA, 37°C, vmax = 280 fmol/min/ng | Escherichia coli | |
| additional information | - |
3-mer 2' phosphorylated RNA | pUpU-(2-phosphate)pU, 37°C, vmax = 1030 fmol/min/ng | Escherichia coli | |
| additional information | - |
[ligated tRNA] | vmax = 0.26 fmol/min/ng | Escherichia coli | |
| 0.0016 | - |
2'-phospho-[ligated tRNA] | - |
Saccharomyces cerevisiae | |
| 0.0057 | - |
3-mer 2' phosphorylated RNA | pApApA-(2-phosphate), 30°C | Saccharomyces cerevisiae | |
| 0.04 | - |
18-mer 2' phosphorylated RNA | pACAAGACUCUAA-(2-phosphate)AUCUUG, 30°C | Saccharomyces cerevisiae | |
| 0.147 | - |
3-mer 2' phosphorylated RNA | pUpU-(2-phosphate)pU, 30°C | Saccharomyces cerevisiae | |
| 0.15 | - |
3-mer 2' phosphorylated RNA | pApApA-(2-phosphate)p, 30°C | Saccharomyces cerevisiae | |
| 0.317 | - |
8-mer 2' phosphorylated RNA | pGUAA-(2-phosphate)AUCU, 30°C | Saccharomyces cerevisiae | |
| 0.45 | - |
3-mer 2' phosphorylated RNA | pApU-(2-phosphate)pA, 30°C | Saccharomyces cerevisiae | |
| 0.73 | - |
3-mer 2' phosphorylated RNA | pApA-(2-phosphate)pA, 30°C | Saccharomyces cerevisiae | |
| 1.173 | - |
3-mer 2' phosphorylated RNA | pApApA-(2-phosphate)pOCH3, 30°C | Saccharomyces cerevisiae | |
| 6.08 | - |
3-mer 2' phosphorylated RNA | pApApA-(2-phosphate)pOCH3, 30°C | Saccharomyces cerevisiae |
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