Any feedback?
Literature summary for 5.6.2.1 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)
| Cloned (Comment) | Organism |
|---|---|
| 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
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| camptothecin | - |
Staphylococcus aureus |  |
| EDTA | complete inhibition of DNA relaxation at 10 mM | Staphylococcus aureus | |
| additional information | no inhibition by nalidixic acid and etoposide, and by spermidine | Staphylococcus aureus | |
| NaCl | inhibits DNA relaxation at over 60 mM | Staphylococcus aureus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Cu2+ | can partially substitute for Mg2+ | Staphylococcus aureus | |
| Mg2+ | dependent on, most favorable divalent cation for the relaxation activity of the enzyme | Staphylococcus aureus | |
| 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 | |
| NaCl | stimulates DNA relaxation at 50 mM | Staphylococcus aureus | |
| Zn2+ | can partially substitute for Mg2+ | Staphylococcus aureus | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Staphylococcus aureus | Q06AK7 | - |
- |
| Staphylococcus aureus C-66 | Q06AK7 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged TopoI from Escherichia coli strain DH5alpha by ammonium sulfate fractionation and nickel affinity chromatography | Staphylococcus aureus |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 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)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 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 | ? | - |
? | |
| 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
| Subunits | Comment | Organism |
|---|---|---|
| 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
| Synonyms | Comment | Organism |
|---|---|---|
| topoI | - |
Staphylococcus aureus |
| type IA DNA topoisomerase | - |
Staphylococcus aureus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | 40 | - |
Staphylococcus aureus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | 70 | 50% of maximal activity at 70°C | Staphylococcus aureus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
- |
Staphylococcus aureus |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 11 | activity range at 37°C, inactive below pH 5.0 | Staphylococcus aureus |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| additional information | no dependence on ATP | Staphylococcus aureus |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
