Literature summary extracted from
Sengupta, T.; Mukherjee, M.; Das, A.; Mandal, C.; Das, R.; Mukherjee, T.; Majumder, H.K.
Characterization of the ATPase activity of topoisomerase II from Leishmania donovani and identification of residues conferring resistance to etoposide (2005), Biochem. J., 390, 419-426.
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
5.6.2.2 |
expression of the His-tagged wild-type and mutant 43 kDa N-terminal domains of the enzyme in Escherichia coli strain BL21(DE3) |
Leishmania donovani |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
5.6.2.2 |
D130G |
site-directed mutagenesis, reduced ATPase activity compared to the wild-type |
Leishmania donovani |
5.6.2.2 |
E155F |
site-directed mutagenesis, mutation of ATP binding pocket residue in the N-terminal domain, no reduction of ATPase activity compared to the wild-type, altered etoposide binding compared the wild-type |
Leishmania donovani |
5.6.2.2 |
G140D |
site-directed mutagenesis, reduced ATPase activity compared to the wild-type |
Leishmania donovani |
5.6.2.2 |
K123A |
site-directed mutagenesis, mutation of ATP binding pocket residue in the N-terminal domain, no reduction of ATPase activity compared to the wild-type, altered etoposide binding compared the wild-type |
Leishmania donovani |
5.6.2.2 |
N65K |
site-directed mutagenesis, mutation of ATP binding pocket residue in the N-terminal domain, no reduction of ATPase activity compared to the wild-type, no inhibition of the mutant by etoposide |
Leishmania donovani |
5.6.2.2 |
N68K |
site-directed mutagenesis, reduced ATPase activity compared to the wild-type |
Leishmania donovani |
5.6.2.2 |
N69K |
site-directed mutagenesis, mutation of ATP binding pocket residue in the N-terminal domain, reduced ATPase activity compared to the wild-type |
Leishmania donovani |
5.6.2.2 |
N96K |
site-directed mutagenesis, mutation of ATP binding pocket residue in the N-terminal domain, no reduction of ATPase activity compared to the wild-type, no inhibition of the mutant by etoposide |
Leishmania donovani |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
5.6.2.2 |
etoposide |
traps the enzyme in a covalent complex with DNA, competes with ATP for the N-terminal binding site, binding involves N65, V77, Y127, N128, and N96, inhibits the ATPase activity of the recombinant N-terminal domain by 73% at 0.01 mM, molecular docking analysis, overview |
Leishmania donovani |
|
5.6.2.2 |
luteolin |
inhibits the ATPase activity of the recombinant N-terminal domain by 57% at 0.02 mM |
Leishmania donovani |
|
5.6.2.2 |
additional information |
no inhibition of ATPase activity by dihydrobetulinic acid, a pentacyclic triterpenoid derivative |
Leishmania donovani |
|
KM Value [mM]
EC Number |
KM Value [mM] |
KM Value Maximum [mM] |
Substrate |
Comment |
Organism |
Structure |
---|
5.6.2.2 |
0.33 |
- |
ATP |
pH 7.5, 30°C, recombinant N-terminal domain, ATPase activity |
Leishmania donovani |
|
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
5.6.2.2 |
KCl |
- |
Leishmania donovani |
|
5.6.2.2 |
Mg2+ |
required by the N-terminal domain for ATPase activity |
Leishmania donovani |
|
5.6.2.2 |
NaCl |
- |
Leishmania donovani |
|
Molecular Weight [Da]
EC Number |
Molecular Weight [Da] |
Molecular Weight Maximum [Da] |
Comment |
Organism |
---|
5.6.2.2 |
43000 |
- |
x * 43000, recombinant N-terminal enzyme domain, SDS-PAGE |
Leishmania donovani |
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
5.6.2.2 |
Leishmania donovani |
Q9XZN0 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
5.6.2.2 |
recombinant His-tagged wild-type and mutant 43 kDa N-terminal domains of the enzyme from Escherichia coli strain BL21(DE3) by nickel affinity and phosphocellulose chromatography |
Leishmania donovani |
Specific Activity [micromol/min/mg]
EC Number |
Specific Activity Minimum [µmol/min/mg] |
Specific Activity Maximum [µmol/min/mg] |
Comment |
Organism |
---|
5.6.2.2 |
additional information |
- |
- |
Leishmania donovani |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
5.6.2.2 |
supercoiled DNA + ATP + H2O |
- |
Leishmania donovani |
catenated DNA networks + ADP + phosphate |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
5.6.2.2 |
? |
x * 43000, recombinant N-terminal enzyme domain, SDS-PAGE |
Leishmania donovani |
5.6.2.2 |
More |
the recombinant 43 kDa N-terminal domain of the enzyme forms a dimer which is stable in absence or presence of nucleotides, structure modeling, overview |
Leishmania donovani |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
5.6.2.2 |
Topoisomerase II |
- |
Leishmania donovani |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
5.6.2.2 |
30 |
- |
ATPase assay at |
Leishmania donovani |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
5.6.2.2 |
7.5 |
- |
ATPase assay at |
Leishmania donovani |
Ki Value [mM]
EC Number |
Ki Value [mM] |
Ki Value maximum [mM] |
Inhibitor |
Comment |
Organism |
Structure |
---|
5.6.2.2 |
0.002 |
- |
etoposide |
pH 7.5, 30°C, recombinant N-terminal domain, inhibition of ATPase activity |
Leishmania donovani |
|