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 | 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Staphylococcus aureus | Q06AK7 | - |
- |
Staphylococcus aureus C-66 | Q06AK7 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged TopoI from Escherichia coli strain DH5alpha by ammonium sulfate fractionation and nickel affinity chromatography | Staphylococcus aureus |
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 | 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 | 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 | Comment | Organism |
---|---|---|
topoI | - |
Staphylococcus aureus |
type IA DNA topoisomerase | - |
Staphylococcus aureus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | 40 | - |
Staphylococcus aureus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | 70 | 50% of maximal activity at 70°C | Staphylococcus aureus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Staphylococcus aureus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 11 | activity range at 37°C, inactive below pH 5.0 | Staphylococcus aureus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | no dependence on ATP | Staphylococcus aureus |