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Literature summary extracted from

  • Garcia, M.T.; Carreno, D.; Tirado-Velez, J.M.; Ferrandiz, M.J.; Rodrigues, L.; Gracia, B.; Amblar, M.; Ainsa, J.A.; de la Campa, A.G.
    Boldine-derived alkaloids inhibit the activity of DNA topoisomerase I and growth of Mycobacterium tuberculosis (2018), Front. Microbiol., 9, 1659.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.6.2.1 gene topA, DNA and amino acid sequence determination and analysis, recombinant expression of His6-tagged enzyme in Escherichia coli strain XL-1 Mycobacterium tuberculosis

Inhibitors

EC Number Inhibitors Comment Organism Structure
5.6.2.1 additional information boldine-derived alkaloids inhibit the activity of DNA topoisomerase I and growth of Mycobacterium tuberculosis Mycobacterium tuberculosis
5.6.2.1 N-methyl-seconeolitsine NSCN, targets DNA topoisomerase I enzyme and inhibits growth of Streptococcus pneumoniae Mycobacterium tuberculosis
5.6.2.1 seconeolitsine SCN, targets DNA topoisomerase I enzyme and inhibits growth of Streptococcus pneumoniae Mycobacterium tuberculosis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
5.6.2.1 100000
-
 about, recombinant His6-tagged enzyme, gel filtration Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
5.6.2.1 Mycobacterium tuberculosis P9WG49
-
-
5.6.2.1 Mycobacterium tuberculosis ATCC 25618 P9WG49
-
-
5.6.2.1 Mycobacterium tuberculosis H37Rv P9WG49
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
5.6.2.1 recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography and dialysis Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.6.2.1 5'-biotin-labeled 32-mer oligonucleotide for the cleavage assay, a 5'-biotin-labeled 32-mer oligonucleotide (5'-CAGTGAGCGAGCTTCCGCTTGACATCCCAATA-3'), which contains the strong topoisomerase site of mycobacterial TopoI, is used as substrate leading to a 19-mer product Mycobacterium tuberculosis ?
-
 ?
5.6.2.1 5'-biotin-labeled 32-mer oligonucleotide for the cleavage assay, a 5'-biotin-labeled 32-mer oligonucleotide (5'-CAGTGAGCGAGCTTCCGCTTGACATCCCAATA-3'), which contains the strong topoisomerase site of mycobacterial TopoI, is used as substrate leading to a 19-mer product Mycobacterium tuberculosis H37Rv ?
-
 ?
5.6.2.1 5'-biotin-labeled 32-mer oligonucleotide for the cleavage assay, a 5'-biotin-labeled 32-mer oligonucleotide (5'-CAGTGAGCGAGCTTCCGCTTGACATCCCAATA-3'), which contains the strong topoisomerase site of mycobacterial TopoI, is used as substrate leading to a 19-mer product Mycobacterium tuberculosis ATCC 25618 ?
-
 ?
5.6.2.1 supercoiled plasmid pBR322 relaxation reaction in vitro Mycobacterium tuberculosis ?
-
 ?
5.6.2.1 supercoiled plasmid pBR322 relaxation reaction in vitro Mycobacterium tuberculosis H37Rv ?
-
 ?
5.6.2.1 supercoiled plasmid pBR322 relaxation reaction in vitro Mycobacterium tuberculosis ATCC 25618 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
5.6.2.1 DNA topoisomerase I
-
 Mycobacterium tuberculosis
5.6.2.1 MtbTopoI
-
 Mycobacterium tuberculosis
5.6.2.1 Rv3646c locus name Mycobacterium tuberculosis
5.6.2.1 TopA
-
 Mycobacterium tuberculosis
5.6.2.1 topoI
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.6.2.1 30
-
 cleavage assay at Mycobacterium tuberculosis
5.6.2.1 37
-
 relaxation assay at Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.6.2.1 8
-
 assay at Mycobacterium tuberculosis

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
5.6.2.1 0.0056
-
 pH 8.0, 37°C, recombinant enzyme Mycobacterium tuberculosis N-methyl-seconeolitsine
5.6.2.1 0.0084
-
 pH 8.0, 37°C, recombinant enzyme Mycobacterium tuberculosis seconeolitsine