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Literature summary extracted from

  • Sengupta, T.; Mukherjee, M.; Das, R.; Das, A.; Majumder, H.K.
    Characterization of the DNA-binding domain and identification of the active site residue in the Gyr A' half of Leishmania donovani topoisomerase II (2005), Nucleic Acids Res., 33, 2364-2373.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.6.2.2 expression of His-tagged wild-type and truncation mutant enzymes in Escherichia coli strain BL21(DE3) Leishmania donovani

Protein Variants

EC Number Protein Variants Comment Organism
5.6.2.2 additional information construction of 4 truncation enzyme mutants, i.e. mutants comprising residues 1-1058, residues 430-1058, residues 430-1236, or residues 430-785, deletion of 178 C-terminal residues remains without effect on the enzyme activity, while deletion of 451 and 429 residues from the C-terminus and the N-terminus, respectively, leads to complete loss of activity, the KCl requirement for mutant activity is altered, and the sensitivity to etoposide is eliminated, all mutants still bind to DNA, overview Leishmania donovani

Inhibitors

EC Number Inhibitors Comment Organism Structure
5.6.2.2 KCl required for DNA cleavage, KCl is inhibitory above 400 mM 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 recombinant 116 kDa mutant, ATP hydrolysis activity, in presence of DNA Leishmania donovani
5.6.2.2 0.55
-
 ATP recombinant 116 kDa mutant, ATP hydrolysis activity, in absence of DNA Leishmania donovani

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
5.6.2.2 Ca2+ divalent cation is required for ATP hydrolysis activity, optimal at 4 mM Leishmania donovani
5.6.2.2 KCl required for DNA cleavage, KCl is inhibitory above 400 mM Leishmania donovani
5.6.2.2 Mg2+ divalent cation is required for ATP hydrolysis activity, optimal at 6 mM Leishmania donovani

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
5.6.2.2 40000
-
 x * 116000, recombinant truncation mutant comprising residues 1-1058, SDS-PAGE, x * 70000, recombinant truncation mutant comprising residues 430-1058, SDS-PAGE, x * 89000, recombinant truncation mutant comprising residues 430-1236, SDS-PAGE, x * 40000, recombinant truncation mutant comprising residues 430-785, SDS-PAGE Leishmania donovani
5.6.2.2 70000
-
 x * 116000, recombinant truncation mutant comprising residues 1-1058, SDS-PAGE, x * 70000, recombinant truncation mutant comprising residues 430-1058, SDS-PAGE, x * 89000, recombinant truncation mutant comprising residues 430-1236, SDS-PAGE, x * 40000, recombinant truncation mutant comprising residues 430-785, SDS-PAGE Leishmania donovani
5.6.2.2 89000
-
 x * 116000, recombinant truncation mutant comprising residues 1-1058, SDS-PAGE, x * 70000, recombinant truncation mutant comprising residues 430-1058, SDS-PAGE, x * 89000, recombinant truncation mutant comprising residues 430-1236, SDS-PAGE, x * 40000, recombinant truncation mutant comprising residues 430-785, SDS-PAGE Leishmania donovani
5.6.2.2 116000
-
 x * 116000, recombinant truncation mutant comprising residues 1-1058, SDS-PAGE, x * 70000, recombinant truncation mutant comprising residues 430-1058, SDS-PAGE, x * 89000, recombinant truncation mutant comprising residues 430-1236, SDS-PAGE, x * 40000, recombinant truncation mutant comprising residues 430-785, SDS-PAGE Leishmania donovani

Organism

EC Number Organism UniProt Comment Textmining
5.6.2.2 Leishmania donovani
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
5.6.2.2 recombinant His-tagged wild-type and truncation mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Leishmania donovani

Reaction

EC Number Reaction Comment Organism Reaction ID
5.6.2.2 ATP-dependent breakage, passage and rejoining of double-stranded DNA active site mapping, catalytic residue Tyr775 in the gyrase half of the Leishmania enzyme is responsible for DNA binding, breakage, and rejoining Leishmania donovani
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.6.2.2 kinetoplast DNA + ATP + H2O cleavage of kinetoplast DNA Leishmania donovani ?
-
 ?
5.6.2.2 additional information ATPase activity is shown by the wild-type enzyme and the truncation mutant comprising residues 1-1058, the enzyme changes the DNA topology by coupling ATP hydrolysis to the transport of one DNA helix through a transient double-stranded break in another Leishmania donovani ?
-
 ?
5.6.2.2 supercoiled pRYG DNA + ATP + H2O relaxation of supercoiled pRYG DNA Leishmania donovani ?
-
 ?

Subunits

EC Number Subunits Comment Organism
5.6.2.2 ? x * 116000, recombinant truncation mutant comprising residues 1-1058, SDS-PAGE, x * 70000, recombinant truncation mutant comprising residues 430-1058, SDS-PAGE, x * 89000, recombinant truncation mutant comprising residues 430-1236, SDS-PAGE, x * 40000, recombinant truncation mutant comprising residues 430-785, SDS-PAGE Leishmania donovani
5.6.2.2 dimer multidomain homodimeric enzyme Leishmania donovani

Synonyms

EC Number Synonyms Comment Organism
5.6.2.2 Topoisomerase II
-
 Leishmania donovani

Cofactor

EC Number Cofactor Comment Organism Structure
5.6.2.2 ATP ATP stimulates dsDNA passage of the wild-type and the 116 kDa mutant enzyme Leishmania donovani