Aarhus University Seal / Aarhus Universitets segl

Real-time analysis of cleavage and religation activity of human topoisomerase 1 based on ternary fluorescence resonance energy transfer DNA substrate

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Standard

Real-time analysis of cleavage and religation activity of human topoisomerase 1 based on ternary fluorescence resonance energy transfer DNA substrate. / Wang, Zhenxing; Ouyang, Hui; Tesauro, Cinzia; Ottaviani, Alessio; He, Yong; Fiorani, Paola; Xie, Hui; Desideri, Alessandro; Fu, Zhifeng.

In: Archives of Biochemistry and Biophysics, Vol. 643, 16.02.2018, p. 1-6.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Harvard

Wang, Z, Ouyang, H, Tesauro, C, Ottaviani, A, He, Y, Fiorani, P, Xie, H, Desideri, A & Fu, Z 2018, 'Real-time analysis of cleavage and religation activity of human topoisomerase 1 based on ternary fluorescence resonance energy transfer DNA substrate', Archives of Biochemistry and Biophysics, vol. 643, pp. 1-6. https://doi.org/10.1016/j.abb.2018.02.006

APA

Wang, Z., Ouyang, H., Tesauro, C., Ottaviani, A., He, Y., Fiorani, P., Xie, H., Desideri, A., & Fu, Z. (2018). Real-time analysis of cleavage and religation activity of human topoisomerase 1 based on ternary fluorescence resonance energy transfer DNA substrate. Archives of Biochemistry and Biophysics, 643, 1-6. https://doi.org/10.1016/j.abb.2018.02.006

CBE

MLA

Vancouver

Author

Wang, Zhenxing ; Ouyang, Hui ; Tesauro, Cinzia ; Ottaviani, Alessio ; He, Yong ; Fiorani, Paola ; Xie, Hui ; Desideri, Alessandro ; Fu, Zhifeng. / Real-time analysis of cleavage and religation activity of human topoisomerase 1 based on ternary fluorescence resonance energy transfer DNA substrate. In: Archives of Biochemistry and Biophysics. 2018 ; Vol. 643. pp. 1-6.

Bibtex

@article{68594bed75284edea1a64bd547c540d2,
title = "Real-time analysis of cleavage and religation activity of human topoisomerase 1 based on ternary fluorescence resonance energy transfer DNA substrate",
abstract = "Human topoisomerase 1B is a ubiquitous and essential enzyme involved in relaxing the topological state of supercoiled DNA to allow the progression of fundamental DNA metabolism. Its enzymatic catalytic cycle consists of cleavage and religation reaction. A ternary fluorescence resonance energy transfer biosensor based on a suicide DNA substrate conjugated with three fluorophores has been developed to monitor both cleavage and religation Topoisomerase I catalytic function. The presence of fluorophores does not alter the specificity of the enzyme catalysis on the DNA substrate. The enzyme-mediated reaction can be tracked in real-time by simple fluorescence measurement, avoiding the use of risky radioactive substrate labeling and time-consuming denaturing gel electrophoresis. The method is applied to monitor the perturbation brought by single mutation on the cleavage or religation reaction and to screen the effect of the camptothecin anticancer drug monitoring the energy transfer decrease during religation reaction. Pathological mutations usually affect only the cleavage or the religation reaction and the proposed approach represent a fast protocol for assessing chemotherapeutic drug efficacy and analyzing mutant's properties.",
keywords = "Enzyme activity, Fluorescence resonance energy transfer, Real-time measurement, Topoisomerase 1",
author = "Zhenxing Wang and Hui Ouyang and Cinzia Tesauro and Alessio Ottaviani and Yong He and Paola Fiorani and Hui Xie and Alessandro Desideri and Zhifeng Fu",
year = "2018",
month = feb,
day = "16",
doi = "10.1016/j.abb.2018.02.006",
language = "English",
volume = "643",
pages = "1--6",
journal = "Nitric Oxide: Biology and Chemistry",
issn = "1089-8603",
publisher = "Academic Press",

}

RIS

TY - JOUR

T1 - Real-time analysis of cleavage and religation activity of human topoisomerase 1 based on ternary fluorescence resonance energy transfer DNA substrate

AU - Wang, Zhenxing

AU - Ouyang, Hui

AU - Tesauro, Cinzia

AU - Ottaviani, Alessio

AU - He, Yong

AU - Fiorani, Paola

AU - Xie, Hui

AU - Desideri, Alessandro

AU - Fu, Zhifeng

PY - 2018/2/16

Y1 - 2018/2/16

N2 - Human topoisomerase 1B is a ubiquitous and essential enzyme involved in relaxing the topological state of supercoiled DNA to allow the progression of fundamental DNA metabolism. Its enzymatic catalytic cycle consists of cleavage and religation reaction. A ternary fluorescence resonance energy transfer biosensor based on a suicide DNA substrate conjugated with three fluorophores has been developed to monitor both cleavage and religation Topoisomerase I catalytic function. The presence of fluorophores does not alter the specificity of the enzyme catalysis on the DNA substrate. The enzyme-mediated reaction can be tracked in real-time by simple fluorescence measurement, avoiding the use of risky radioactive substrate labeling and time-consuming denaturing gel electrophoresis. The method is applied to monitor the perturbation brought by single mutation on the cleavage or religation reaction and to screen the effect of the camptothecin anticancer drug monitoring the energy transfer decrease during religation reaction. Pathological mutations usually affect only the cleavage or the religation reaction and the proposed approach represent a fast protocol for assessing chemotherapeutic drug efficacy and analyzing mutant's properties.

AB - Human topoisomerase 1B is a ubiquitous and essential enzyme involved in relaxing the topological state of supercoiled DNA to allow the progression of fundamental DNA metabolism. Its enzymatic catalytic cycle consists of cleavage and religation reaction. A ternary fluorescence resonance energy transfer biosensor based on a suicide DNA substrate conjugated with three fluorophores has been developed to monitor both cleavage and religation Topoisomerase I catalytic function. The presence of fluorophores does not alter the specificity of the enzyme catalysis on the DNA substrate. The enzyme-mediated reaction can be tracked in real-time by simple fluorescence measurement, avoiding the use of risky radioactive substrate labeling and time-consuming denaturing gel electrophoresis. The method is applied to monitor the perturbation brought by single mutation on the cleavage or religation reaction and to screen the effect of the camptothecin anticancer drug monitoring the energy transfer decrease during religation reaction. Pathological mutations usually affect only the cleavage or the religation reaction and the proposed approach represent a fast protocol for assessing chemotherapeutic drug efficacy and analyzing mutant's properties.

KW - Enzyme activity

KW - Fluorescence resonance energy transfer

KW - Real-time measurement

KW - Topoisomerase 1

UR - http://www.scopus.com/inward/record.url?scp=85042237742&partnerID=8YFLogxK

U2 - 10.1016/j.abb.2018.02.006

DO - 10.1016/j.abb.2018.02.006

M3 - Journal article

C2 - 29458004

AN - SCOPUS:85042237742

VL - 643

SP - 1

EP - 6

JO - Nitric Oxide: Biology and Chemistry

JF - Nitric Oxide: Biology and Chemistry

SN - 1089-8603

ER -