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Literature summary for 3.2.2.27 extracted from
- Cloning, expression, and characterization of uracil-DNA glycosylase of Chlamydia pneumoniae in Escherichia coli (2004), Protein Expr. Purif., 35, 46-53.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of His-tagged wild-type and mutant UDGs in Escherichia coli strain BL21(DE3) | Chlamydia pneumoniae |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A205S | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Chlamydia pneumoniae |
| D77N | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Chlamydia pneumoniae |
| H200Q | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Chlamydia pneumoniae |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| additional information | free uracil and abasic sites inhibit the enzyme reaction | Chlamydia pneumoniae |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0052 | - |
uracil-mismatched double-stranded DNA | pH 8.0, 37°C, recombinant wild-type enzyme | Chlamydia pneumoniae | |
| 0.009 | - |
uracil-mismatched double-stranded DNA | pH 8.0, 37°C, recombinant mutant A205S | Chlamydia pneumoniae | |
| 0.148 | - |
uracil-mismatched double-stranded DNA | pH 8.0, 37°C, recombinant mutant H200Q | Chlamydia pneumoniae | |
| 0.256 | - |
uracil-mismatched double-stranded DNA | pH 8.0, 37°C, recombinant mutant D77N | Chlamydia pneumoniae |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Chlamydia pneumoniae | Q9Z7D3 | - |
- |
| Chlamydia pneumoniae AR39 | Q9Z7D3 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutant UDGs from Escherichia coli strain BL21 (DE3) by nickel affinity and anion exchange chromatography | Chlamydia pneumoniae |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
purified recombinant enzyme | Chlamydia pneumoniae |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| uracil-mismatched double-stranded DNA + H2O | duplex DNA with A/U mismatch, substrate specificity, the base opposite to uracil in double strand DNAs affects uracil removal efficiencies in descending order U/-, U/T, U/C, U/G, U/A. Free uracil and abasic sites inhibit the reaction. Amino acids D77, H200, and A205 are important for the catalytic activity of UDG | Chlamydia pneumoniae | uracil + double-stranded DNA with abasic site | - |
? |  |
| uracil-mismatched double-stranded DNA + H2O | duplex DNA with A/U mismatch, substrate specificity, the base opposite to uracil in double strand DNAs affects uracil removal efficiencies in descending order U/-, U/T, U/C, U/G, U/A. Free uracil and abasic sites inhibit the reaction. Amino acids D77, H200, and A205 are important for the catalytic activity of UDG | Chlamydia pneumoniae AR39 | uracil + double-stranded DNA with abasic site | - |
? | |
| uracil-mismatched double-stranded DNA with A-U mismatch + H2O | - |
Chlamydia pneumoniae | uracil + double-stranded DNA with abasic site | - |
? | |
| uracil-mismatched double-stranded DNA with A-U mismatch + H2O | - |
Chlamydia pneumoniae AR39 | uracil + double-stranded DNA with abasic site | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| More | the enzyme is a member of the UDG family I | Chlamydia pneumoniae |
| UDG | - |
Chlamydia pneumoniae |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
- |
Chlamydia pneumoniae |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.042 | - |
uracil-mismatched double-stranded DNA | pH 8.0, 37°C, recombinant mutant D77N | Chlamydia pneumoniae | |
| 0.053 | - |
uracil-mismatched double-stranded DNA | pH 8.0, 37°C, recombinant mutant H200Q | Chlamydia pneumoniae | |
| 0.73 | - |
uracil-mismatched double-stranded DNA | pH 8.0, 37°C, recombinant mutant A205S | Chlamydia pneumoniae | |
| 5.9 | - |
uracil-mismatched double-stranded DNA with A-U mismatch + | pH 8.0, 37°C, recombinant wild-type enzyme | Chlamydia pneumoniae |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
- |
Chlamydia pneumoniae |
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