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Literature summary extracted from
- Cloning and biochemical characterization of Staphylococcus aureus type IA DNA topoisomerase comprised of distinct five domains (2011), Arch. Biochem. Biophys., 508, 78-86.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 5.6.2.1 | gene Sau-TopoI, DNA and amino acid sequence determination and analysis, sequence comparisons, expression of His-tagged enzyme in Escherichia coli strain DH5alpha | Staphylococcus aureus |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.6.2.1 | camptothecin | - |
Staphylococcus aureus |  |
| 5.6.2.1 | EDTA | complete inhibition of DNA relaxation at 10 mM | Staphylococcus aureus | |
| 5.6.2.1 | additional information | no inhibition by nalidixic acid and etoposide, and by spermidine | Staphylococcus aureus | |
| 5.6.2.1 | NaCl | inhibits DNA relaxation at over 60 mM | Staphylococcus aureus | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.6.2.1 | Cu2+ | can partially substitute for Mg2+ | Staphylococcus aureus | |
| 5.6.2.1 | Mg2+ | dependent on, most favorable divalent cation for the relaxation activity of the enzyme | Staphylococcus aureus | |
| 5.6.2.1 | additional information | the enzyme shows no relaxation activity on supercoiled DNA in the absence of divalent cations, no or poor stimulation by Ca2+ and Mn2+ | Staphylococcus aureus | |
| 5.6.2.1 | NaCl | stimulates DNA relaxation at 50 mM | Staphylococcus aureus | |
| 5.6.2.1 | Zn2+ | can partially substitute for Mg2+ | Staphylococcus aureus | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 5.6.2.1 | Staphylococcus aureus | Q06AK7 | - |
- |
| 5.6.2.1 | Staphylococcus aureus C-66 | Q06AK7 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 5.6.2.1 | recombinant His-tagged TopoI from Escherichia coli strain DH5alpha by ammonium sulfate fractionation and nickel affinity chromatography | Staphylococcus aureus |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 5.6.2.1 | additional information | - |
5000 U, with one unit topoisomerase activity defined as the amount of enzyme required to relax 50% of supercoiled pGEM3zf(-) DNA (200 ng) for 30 min at 37°C, pH 8.0, in the presence of 5 mM of MgCl2, recombinant enzyme | Staphylococcus aureus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.6.2.1 | additional information | Sau-TopoI enzyme converts negatively supercoiled form I DNA to relaxed form II DNA in ATP-independent and Mg2+-dependent manners. Cleavage site mapping shows that the enzyme preferentially binds to and cleaves the sequence GGNN-/-CAT, with N and -/- representing any nucleotide and cleavage site, respectively | Staphylococcus aureus | ? | - |
? | |
| 5.6.2.1 | additional information | Sau-TopoI enzyme converts negatively supercoiled form I DNA to relaxed form II DNA in ATP-independent and Mg2+-dependent manners. Cleavage site mapping shows that the enzyme preferentially binds to and cleaves the sequence GGNN-/-CAT, with N and -/- representing any nucleotide and cleavage site, respectively | Staphylococcus aureus C-66 | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 5.6.2.1 | More | domain mapping analysis shows that the enzyme is composed of conserved four domains, I through IV, together with a variable C-terminal region containing a unique domain V | Staphylococcus aureus |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 5.6.2.1 | topoI | - |
Staphylococcus aureus |
| 5.6.2.1 | type IA DNA topoisomerase | - |
Staphylococcus aureus |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 5.6.2.1 | 37 | 40 | - |
Staphylococcus aureus |
Temperature Range [°C]
| EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 5.6.2.1 | 37 | 70 | 50% of maximal activity at 70°C | Staphylococcus aureus |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.6.2.1 | 7.5 | - |
- |
Staphylococcus aureus |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.6.2.1 | 6 | 11 | activity range at 37°C, inactive below pH 5.0 | Staphylococcus aureus |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.6.2.1 | additional information | no dependence on ATP | Staphylococcus aureus |
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