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AB82099

Recombinant Human Topoisomerase I protein

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Recombinant Human Topoisomerase I protein is a Human Full Length protein, in the 1 to 765 aa range, expressed in Baculovirus, with >95%, suitable for SDS-PAGE.

View Alternative Names

DNA topoisomerase 1, DNA topoisomerase I, TOP1

Key facts

Purity

>95% SDS-PAGE

Expression system

Baculovirus

Tags

6x His tag N-Terminus

Applications

SDS-PAGE

applications

Biologically active

No

Accession

P11387

Animal free

No

Carrier free

No

Species

Human

Storage buffer

pH: 7.9 Constituents: 20% Glycerol (glycerin, glycerine), 0.75% Potassium chloride, 0.316% Tris HCl, 0.0154% (R*,R*)-1,4-Dimercaptobutan-2,3-diol, 0.00584% EDTA

storage-buffer

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "SDS-PAGE": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

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.

Purified topo I has been used for in vitro transcriptional activation, pre-mRNA splicing/phosphorylation, DNA binding and DNA relaxation assays.

Sequence info

[{"sequence":"MSGDHLHNDSQIEADFRLNDSHKHKDKHKDREHRHKEHKKEKDREKSKHSNSEHKDSEKKHKEKEKTKHKDGSSEKHKDKHKDRDKEKRKEEKVRASGDAKIKKEKENGFSSPPQIKDEPEDDGYFVPPKEDIKPLKRPRDEDDADYKPKKIKTEDTKKEKKRKLEEEEDGKLKKPKNKDKDKKVPEPDNKKKKPKKEEEQKWKWWEEERYPEGIKWKFLEHKGPVFAPPYEPLPENVKFYYDGKVMKLSPKAEEVATFFAKMLDHEYTTKEIFRKNFFKDWRKEMTNEEKNIITNLSKCDFTQMSQYFKAQTEARKQMSKEEKLKIKEENEKLLKEYGFCIMDNHKERIANFKIEPPGLFRGRGNHPKMGMLKRRIMPEDIIINCSKDAKVPSPPPGHKWKEVRHDNKVTWLVSWTENIQGSIKYIMLNPSSRIKGEKDWQKYETARRLKKCVDKIRNQYREDWKSKEMKVRQRAVALYFIDKLALRAGNEKEEGETADTVGCCSLRVEHINLHPELDGQEYVVEFDFLGKDSIRYYNKVPVEKRVFKNLQLFMENKQPEDDLFDRLNTGILNKHLQDLMEGLTAKVFRTYNASITLQQQLKELTAPDENIPAKILSYNRANRAVAILCNHQRAPPKTFEKSMMNLQTKIDAKKEQLADARRDLKSAKADAKVMKDAKTKKVVESKKKAVQRLEEQLMKLEVQATDREENKQIALGTSKLNYLDPRITVAWCKKWGVPIEKIYNKTQREKFAWAIDMADEDYEF","proteinLength":"Full Length","predictedMolecularWeight":null,"actualMolecularWeight":null,"aminoAcidEnd":765,"aminoAcidStart":1,"nature":"Recombinant","expressionSystem":"Baculovirus","accessionNumber":"P11387","tags":[{"tag":"6x His","terminus":"N-Terminus"}]}]

Properties and storage information

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-80°C
Appropriate long-term storage conditions
-80°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle
False

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Topoisomerase I also known as DNA topoisomerase I or Topo I is an important enzyme in the unwinding of the DNA double helix during processes such as replication and transcription. This enzyme has a molecular mass of approximately 91 kDa and functions by creating transient single-strand breaks in the DNA allowing relaxation of supercoils. Topoisomerase I is widely expressed in both proliferating and non-proliferating cells with higher levels seen in tissues with rapid cell division like the gastrointestinal tract and some immune cells.
Biological function summary

Topoisomerase I plays a significant role in the modulation of DNA topology ensuring proper chromosomal functions during cellular proliferation. It serves as a single polypeptide and does not require a companion for its activity unlike other topoisomerases that may function in complexes. The enzyme’s ability to relieve torsional stress in DNA is essential for maintaining genomic stability and facilitating the smooth progression of the replication fork.

Pathways

Topoisomerase I is vital for DNA replication and transcription processes. It works in concert with other proteins such as helicases and ligases to ensure efficient unwinding and rewinding of DNA strands. The enzyme participates in the DNA damage response pathway where it interacts with proteins like PARP1 to coordinate repair processes. Its action is important in both the S phase of the cell cycle where DNA synthesis occurs and in the G0 phase where cells are in a quiescent state.

Topoisomerase I is heavily implicated in oncogenesis with significant associations to colorectal and ovarian cancers. Its heightened activity can lead to genomic instability a hallmark of cancer development. The enzyme also relates to chemotherapeutic resistance where mutations in topoisomerase I lead to reduced drug efficacy. Furthermore inhibitors targeting topoisomerase I such as camptothecin-based drugs exploit its role in cancer aiming to induce DNA damage selectively in rapidly dividing cells. Connections to other proteins like BRCA1 are apparent in these pathways as they both contribute to the DNA repair processes in cells.

Specifications

Form

Liquid

Additional notes

Purified by using an affinity column and FPLC chromatography.

General info

Function

Releases the supercoiling and torsional tension of DNA introduced during the DNA replication and transcription by transiently cleaving and rejoining one strand of the DNA duplex. Introduces a single-strand break via transesterification at a target site in duplex DNA. The scissile phosphodiester is attacked by the catalytic tyrosine of the enzyme, resulting in the formation of a DNA-(3'-phosphotyrosyl)-enzyme intermediate and the expulsion of a 5'-OH DNA strand. The free DNA strand then rotates around the intact phosphodiester bond on the opposing strand, thus removing DNA supercoils. Finally, in the religation step, the DNA 5'-OH attacks the covalent intermediate to expel the active-site tyrosine and restore the DNA phosphodiester backbone (By similarity). Regulates the alternative splicing of tissue factor (F3) pre-mRNA in endothelial cells. Involved in the circadian transcription of the core circadian clock component BMAL1 by altering the chromatin structure around the ROR response elements (ROREs) on the BMAL1 promoter.

Sequence similarities

Belongs to the type IB topoisomerase family.

Post-translational modifications

Sumoylated. Lys-117 is the main site of sumoylation. Sumoylation plays a role in partitioning TOP1 between nucleoli and nucleoplasm. Levels are dramatically increased on camptothecin (CPT) treatment.. Phosphorylation at Ser-506 by CK2 increases binding to supercoiled DNA and sensitivity to camptothecin.

Subcellular localisation

Nucleus

Product protocols

Target data

Releases the supercoiling and torsional tension of DNA introduced during the DNA replication and transcription by transiently cleaving and rejoining one strand of the DNA duplex. Introduces a single-strand break via transesterification at a target site in duplex DNA. The scissile phosphodiester is attacked by the catalytic tyrosine of the enzyme, resulting in the formation of a DNA-(3'-phosphotyrosyl)-enzyme intermediate and the expulsion of a 5'-OH DNA strand. The free DNA strand then rotates around the intact phosphodiester bond on the opposing strand, thus removing DNA supercoils. Finally, in the religation step, the DNA 5'-OH attacks the covalent intermediate to expel the active-site tyrosine and restore the DNA phosphodiester backbone (By similarity). Regulates the alternative splicing of tissue factor (F3) pre-mRNA in endothelial cells. Involved in the circadian transcription of the core circadian clock component BMAL1 by altering the chromatin structure around the ROR response elements (ROREs) on the BMAL1 promoter.
See full target information TOP1

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