A fluorescence based assay for the AP-site cleavage activity of human tyrosyl DNA phosphodiesterase 1

Graeme J. Thomson*; Nicola S. Hamilton; Gemma V. Hopkins; Ian D. Waddell; Amanda J. Watson; Donald J. Ogilvie

A fluorescence based assay for the AP-site cleavage activity of human tyrosyl DNA phosphodiesterase 1

Graeme J. Thomson*, Nicola S. Hamilton, Gemma V. Hopkins, Ian D. Waddell, Amanda J. Watson, Donald J. Ogilvie

Cancer Research UK Manchester Institute

Research output: Contribution to journal › Article › peer-review

Abstract

Tyrosyl-DNA phosphodiesterase 1 (Tdp1) catalyzes the hydrolysis of phosphodiester bonds between the DNA 30-phosphate and tyrosine residues and plays a major role in the repair of stalled topoisomerase IDNA covalent complexes. Given this role, Tdp1 is of interest as a potential target for anticancer therapy. Inhibiting Tdp1 in combination with clinically used Top1 inhibitors may potentiate the effects of the latter and help to overcome some of the chemoresistance issues currently observed. In addition, Tdp1 can function during DNA repair to remove a variety of other 30 adducts from DNA such as phosphoglycolates and abasic or apurinic/apyrimidinic (AP) sites. Here we describe a new mix-and-read homogeneous fluorogenic assay for the measurement of the AP-site cleavage activity of Tdp1 that is compatible with highthroughput screening. The application of such an assay will open up further avenues for the discovery of novel Tdp1 inhibitors.

Original language	English
Pages (from-to)	1-5
Number of pages	4
Journal	Analytical Biochemistry
Volume	440
Publication status	Published - 2013

Keywords

TDP1

Research Beacons, Institutes and Platforms

Manchester Cancer Research Centre

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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@article{16f3f02a0eee419b83b63412ba100cbc,

title = "A fluorescence based assay for the AP-site cleavage activity of human tyrosyl DNA phosphodiesterase 1",

abstract = "Tyrosyl-DNA phosphodiesterase 1 (Tdp1) catalyzes the hydrolysis of phosphodiester bonds between the DNA 30-phosphate and tyrosine residues and plays a major role in the repair of stalled topoisomerase IDNA covalent complexes. Given this role, Tdp1 is of interest as a potential target for anticancer therapy. Inhibiting Tdp1 in combination with clinically used Top1 inhibitors may potentiate the effects of the latter and help to overcome some of the chemoresistance issues currently observed. In addition, Tdp1 can function during DNA repair to remove a variety of other 30 adducts from DNA such as phosphoglycolates and abasic or apurinic/apyrimidinic (AP) sites. Here we describe a new mix-and-read homogeneous fluorogenic assay for the measurement of the AP-site cleavage activity of Tdp1 that is compatible with highthroughput screening. The application of such an assay will open up further avenues for the discovery of novel Tdp1 inhibitors.",

keywords = "TDP1",

author = "Thomson*, {Graeme J.} and Hamilton, {Nicola S.} and Hopkins, {Gemma V.} and Waddell, {Ian D.} and Watson, {Amanda J.} and Ogilvie, {Donald J.}",

note = "This work was supported by Cancer Research UK (C480/ A11411).",

year = "2013",

language = "English",

volume = "440",

pages = "1--5",

journal = "Analytical Biochemistry",

issn = "0003-2697",

publisher = "Academic Press, Ltd",

}

TY - JOUR

T1 - A fluorescence based assay for the AP-site cleavage activity of human tyrosyl DNA phosphodiesterase 1

AU - Thomson, Graeme J.

AU - Hamilton, Nicola S.

AU - Hopkins, Gemma V.

AU - Waddell, Ian D.

AU - Watson, Amanda J.

AU - Ogilvie, Donald J.

N1 - This work was supported by Cancer Research UK (C480/ A11411).

PY - 2013

Y1 - 2013

N2 - Tyrosyl-DNA phosphodiesterase 1 (Tdp1) catalyzes the hydrolysis of phosphodiester bonds between the DNA 30-phosphate and tyrosine residues and plays a major role in the repair of stalled topoisomerase IDNA covalent complexes. Given this role, Tdp1 is of interest as a potential target for anticancer therapy. Inhibiting Tdp1 in combination with clinically used Top1 inhibitors may potentiate the effects of the latter and help to overcome some of the chemoresistance issues currently observed. In addition, Tdp1 can function during DNA repair to remove a variety of other 30 adducts from DNA such as phosphoglycolates and abasic or apurinic/apyrimidinic (AP) sites. Here we describe a new mix-and-read homogeneous fluorogenic assay for the measurement of the AP-site cleavage activity of Tdp1 that is compatible with highthroughput screening. The application of such an assay will open up further avenues for the discovery of novel Tdp1 inhibitors.

AB - Tyrosyl-DNA phosphodiesterase 1 (Tdp1) catalyzes the hydrolysis of phosphodiester bonds between the DNA 30-phosphate and tyrosine residues and plays a major role in the repair of stalled topoisomerase IDNA covalent complexes. Given this role, Tdp1 is of interest as a potential target for anticancer therapy. Inhibiting Tdp1 in combination with clinically used Top1 inhibitors may potentiate the effects of the latter and help to overcome some of the chemoresistance issues currently observed. In addition, Tdp1 can function during DNA repair to remove a variety of other 30 adducts from DNA such as phosphoglycolates and abasic or apurinic/apyrimidinic (AP) sites. Here we describe a new mix-and-read homogeneous fluorogenic assay for the measurement of the AP-site cleavage activity of Tdp1 that is compatible with highthroughput screening. The application of such an assay will open up further avenues for the discovery of novel Tdp1 inhibitors.

KW - TDP1

M3 - Article

SN - 0003-2697

VL - 440

SP - 1

EP - 5

JO - Analytical Biochemistry

JF - Analytical Biochemistry

ER -

A fluorescence based assay for the AP-site cleavage activity of human tyrosyl DNA phosphodiesterase 1

Abstract

Keywords

Research Beacons, Institutes and Platforms

UN SDGs

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