Recombinant Human Topoisomerase (DNA) I

• Cat.No.: TOP1-519H
• Product Overview: Recombinant Human Topoisomerase (DNA) I protein (765 amino acids) was expressed inBaculovirussystem.

Specification

• Species: Human
• Source: Insect Cells
• Tag: Non
• Description: Human DNA topoisomerase I (topo I) is a monomeric protein of 765 anino acids encoded by a single-copy gene. Based on its function, DNA topoisomerase I has been subdivided into four distinct domains. The N-terminal 214 amino acids of topo I comprise a highly charged N-terminal domain involved in protein-protein interactions with a number of cellular proteins. The highly conserved core domain expanded from amino acids 215 to 636 contains most catalytic residues and retains DNA binding activity. The active C-terminal domain (residues 713-765) is connected to the core domain by a poorly conserved linker domain (residues 636-712).
• Purity: Purity > 95% bySDS-PAGE analysis.
• Endotoxin: 1 unit equals 1 nanogram purified protein. 0.1 – 1 unit (ng) is sufficient for relaxing 0.5 µg of pBR322 supercoiled DNA in a 20 μl reaction.

Gene Information

• Gene Name: TOP1 topoisomerase (DNA) I [ Homo sapiens ]
• Synonyms: TOP1; topoisomerase (DNA) I; TOPI; DNA topoisomerase1; EC 5.99.1.2; OTTHUMP00000031713; DNA topoisomerase I; topoisomerase I; type I DNA topoisomerase
• Gene ID: 7150
• mRNA Refseq: NM_003286
• Protein Refseq: NP_003277
• MIM: 126420
• UniProt ID: P11387
• Chromosome Location: 20q12-q13.1
• Function: ATP binding; DNA topoisomerase (ATP-hydrolyzing) activity; DNA topoisomerase type I activity; chromatin binding; nucleotide binding; protein binding

Citation

Suppression of lupus nephritis and skin lesions in MRL/ lpr mice by administration of the topoisomerase I inhibitor irinotecan

Journal: Arthritis Research & Therapy    PubMed ID: 27770825    Data: 2016/10/22

Authors: Andreas Keil, Sean R. Hall, Steffen Frese

Article Snippet:Calf thymus DNA (50 mg/ml) was incubated with 4 μg/ml recombinant human topo I (Creative BioMart) in 50 mM Tris HCl (pH 7.5), 50 mM KCl, 2 mM dithiothreitol (DTT), and 1 mM EDTA for 30 min at 37 °C.. Then, 30 ml of sample per well was used for coating a 384-well Nunc Maxi-Sorp plate overnight at 4 °C.Then, 30 ml of sample per well was used for coating a 384-well Nunc Maxi-Sorp plate overnight at 4 °C.

Increased binding of anti-double-stranded DNS ( dsDNA ) IgG but not of anti-dsDNA IgM to DNA treated with topoisomerase I ( topo I ). Calf thymus DNA was incubated with recombinant topo I or bovine serum albumin ( BSA ) for 30 min. Binding affinity of a anti-dsDNA IgG or b anti-dsDNA IgM from MRL/ lpr plasma to modified DNA was assessed by enzyme-linked immunosorbent assay. Data were pooled from three independent experiments. ** P < 0.01, by paired t test. ns not significant, OD optical density

The Topoisomerase I Inhibitor Irinotecan and the Tyrosyl-DNA Phosphodiesterase 1 Inhibitor Furamidine Synergistically Suppress Murine Lupus Nephritis.

Journal: Arthritis & rheumatology (Hoboken, N.J.)    PubMed ID: 25779651    Data: 2015/9/16

Authors: Andreas Keil, Manuela Frese-Schaper, Steffen Frese

Article Snippet:Bound anti-dsDNA autoantibodies were detected as described above for total IgG ELISA.Bound anti-dsDNA autoantibodies were detected as described above for total IgG ELISA.. Effect of DNA modification on anti-dsDNA binding.. Calf thymus DNA (50 mg/ml) was incubated with 3.75 mg/ml recombinant human topo I (Creative BioMart) in 50 mM Tris HCl (pH 7.5), 50 mM KCl, 2 mM dithiothreitol (DTT), 1 mM EDTA, and 50% glycerol for 30 minutes at 378C.. Then, 30 ml of sample per well was used for coating a 384-well Nunc MaxiSorp plate overnight at 48C.Then, 30 ml of sample per well was used for coating a 384-well Nunc MaxiSorp plate overnight at 48C.

Low-dose irinotecan improves advanced lupus nephritis in mice potentially by changing DNA relaxation and anti-double-stranded DNA binding.

Journal: Arthritis & rheumatology (Hoboken, N.J.)    PubMed ID: 24729466    Data: 2014/9/22

Authors: Manuela Frese-Schaper, Andreas Keil, Steffen Frese

Article Snippet:Modification of DNA using topo I alone or in combination with the topo I inhibitor camptothecin was performed essentially as previously described (22).Modification of DNA using topo I alone or in combination with the topo I inhibitor camptothecin was performed essentially as previously described (22).. Fifty micrograms of filtered calf thymus dsDNA, nucleosomal DNA, or AT-rich dsDNA poly(dA–dT) (Sigma) was incubated with different concentrations of recombinant human topo I (Creative BioMart) using 0.9 ng, 2.8 ng, 8.3 ng, 25 ng, 75 ng, or 225 ng per g DNA in a 1 ml reaction containing 40 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2, 0.5 mM dithiothreitol, 0.5 mM EDTA, and 30 g/ml bovine serum albumin (BSA; Sigma) for 30 minutes at 37°C.. Then, 384-well Nunc MaxiSorp plates were coated with 30 l per well overnight at 4°C.Then, 384-well Nunc MaxiSorp plates were coated with 30 l per well overnight at 4°C.