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Literature summary for 5.6.2.1 extracted from

  • Godbole, A.A.; Leelaram, M.N.; Bhat, A.G.; Jain, P.; Nagaraja, V.
    Characterization of DNA topoisomerase I from Mycobacterium tuberculosis: DNA cleavage and religation properties and inhibition of its activity (2012), Arch. Biochem. Biophys., 528, 197-203.
    View publication on PubMed
Search for more literature for 5.6.2.1

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of wild-type and mutant His-tagged and non-tagged enzymes in Escherichia coli strain BL21(DE3) Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
D111A site-directed mutagenesis, mutation of the Mg2+-binding residue affects DNA relaxation activity, mutant D111A is dependent on Mg2+ for DNA cleavage and is compromised in religation Mycobacterium tuberculosis
D113A site-directed mutagenesis Mycobacterium tuberculosis
E115A site-directed mutagenesis, mutation of the Mg2+-binding residue affects DNA relaxation activity, the mutant is compromised in religation Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
additional information oligonucleotides containing the specific recognition sequence inhibit the activity of the enzyme, as well as the monoclonal antibody 2F3G4, developed against MstopoI inhibited the relaxation activity of the enzyme. No inhibition with non-specific oligonucleotides Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required, acidic residues D111 and E115 are involved in Mg2+ coordination Mycobacterium tuberculosis

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis A5U8X0
-
-
Mycobacterium tuberculosis H37Ra / ATCC 25177 A5U8X0
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant nontagged enzyme from Escherichia coli strain BL21 by ammonium sulfate fractionation, dialysis, and heparin affinity chromatography, recombinant wild-type and mutant His-tagged enzymes from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, dialysis, and nickel affinity chromatography, native enzyme from Mycobacterium tuberculosis strain H37Ra Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information enzyme activity on relaxation of supercoiled pUC18 DNA or on 5' end labeled oligonucleotides having the strong topoisomerase I site, the enzyme cleaves DNA at preferred sites in a pattern similar to its ortholog from Mycobacterium smegmatis Mycobacterium tuberculosis ?
-
 ?
additional information enzyme activity on relaxation of supercoiled pUC18 DNA or on 5' end labeled oligonucleotides having the strong topoisomerase I site, the enzyme cleaves DNA at preferred sites in a pattern similar to its ortholog from Mycobacterium smegmatis Mycobacterium tuberculosis H37Ra / ATCC 25177 ?
-
 ?

Synonyms

Synonyms Comment Organism
DNA topoisomerase I
-
 Mycobacterium tuberculosis
MttopoI
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Mycobacterium tuberculosis