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Literature summary extracted from
- Pseudouridine synthetase Pus1 of Saccharomyces cerevisiae: kinetic characterisation, tRNA structural requirement and real-time analysis of its complex with tRNA (1999), J. Mol. Biol., 289, 491-502.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 5.4.99.B22 | recombinant Pus1, tagged with six histidine residues at its N-terminus is expressed in Escherichia coli | Saccharomyces cerevisiae |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.4.99.B22 | EDTA | the complex Pus1/tRNAVal is sensitive to the presence of EDTA | Saccharomyces cerevisiae |  |
| 5.4.99.B22 | KCl | the complex of Pus1 with tRNA is sensitive to high ionic strength. The optimal concentration of KCl for Pus1 binding is 100 mM. Above 400 mM KCl, no complex is detected | Saccharomyces cerevisiae | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 4.2.1.70 | 0.00042 | - |
tRNAVal | Pus1, yeast tRNAVal | Saccharomyces cerevisiae | |
| 4.2.1.70 | 0.00074 | - |
tRNAIle | Pus1, intronless yeast tRNAIle | Saccharomyces cerevisiae | |
| 4.2.1.70 | 0.0015 | - |
tRNAVal | Pus1, yeast tRNAVal, in the presence of 5 mM EDTA and in the absence of Mg2+ | Saccharomyces cerevisiae | |
| 5.4.99.B22 | 0.00042 | - |
tRNAVal uridine27 | pH and temperature not specified in the publication | Saccharomyces cerevisiae | |
| 5.4.99.B22 | 0.00074 | - |
tRNAIle uridine27 | pH and temperature not specified in the publication | Saccharomyces cerevisiae |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.4.99.B22 | KCl | the complex of Pus1 with tRNA is sensitive to high ionic strength. The optimal concentration of KCl for Pus1 binding is 100 mM. Above 400 mM KCl, no complex is detected | Saccharomyces cerevisiae | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 4.2.1.70 | additional information | Saccharomyces cerevisiae | pseudouridine synthase Pus1p displays a multisite substrate specificity | ? | - |
? | |
| 4.2.1.70 | additional information | Saccharomyces cerevisiae | a model for Pus1 mechanism | ? | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 4.2.1.70 | Saccharomyces cerevisiae | - |
- |
- |
| 5.4.99.B22 | Saccharomyces cerevisiae | Q12211 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 5.4.99.B22 | recombinant enzyme | Saccharomyces cerevisiae |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 4.2.1.70 | additional information | - |
- |
Saccharomyces cerevisiae |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 4.2.1.70 | additional information | pseudouridine synthase Pus1p displays a multisite substrate specificity | Saccharomyces cerevisiae | ? | - |
? | |
| 4.2.1.70 | additional information | a model for Pus1 mechanism | Saccharomyces cerevisiae | ? | - |
? | |
| 4.2.1.70 | tRNAIle | - |
Saccharomyces cerevisiae | ? | - |
? | |
| 4.2.1.70 | tRNAVal | - |
Saccharomyces cerevisiae | ? | - |
? | |
| 5.4.99.B22 | additional information | Pus1 from Saccharomyces cerevisiae is a multisite-specific enzyme that catalyses the formation of pseudouridine residues at different positions in several tRNA transcripts | Saccharomyces cerevisiae | ? | - |
? | |
| 5.4.99.B22 | tRNAIle uridine27 | - |
Saccharomyces cerevisiae | tRNAIle pseudouridine27 | - |
? | |
| 5.4.99.B22 | tRNAVal uridine27 | - |
Saccharomyces cerevisiae | tRNAVal pseudouridine27 | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 5.4.99.B22 | PUS1 | - |
Saccharomyces cerevisiae |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 4.2.1.70 | 0.00667 | - |
tRNAVal | substrate from yeast, limiting | Saccharomyces cerevisiae | |
| 4.2.1.70 | 0.00833 | - |
tRNAVal | substrate from yeast, in absence of Mg2+ | Saccharomyces cerevisiae | |
| 4.2.1.70 | 0.00833 | - |
tRNAIle | substrate from yeast, intronless, limiting | Saccharomyces cerevisiae | |
| 5.4.99.B22 | 0.0067 | - |
tRNAVal uridine27 | pH and temperature not specified in the publication | Saccharomyces cerevisiae | |
| 5.4.99.B22 | 0.0083 | - |
tRNAIle uridine27 | pH and temperature not specified in the publication | Saccharomyces cerevisiae |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.4.99.B22 | 11.26 | - |
tRNAIle uridine27 | pH and temperature not specified in the publication | Saccharomyces cerevisiae | |
| 5.4.99.B22 | 15.95 | - |
tRNAVal uridine27 | pH and temperature not specified in the publication | Saccharomyces cerevisiae |
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