The establishment of methods for free PAR generation and PAR reader detection

Yueshuang Ke; Ke Wang; Hui Xu; Chenxin Wang; Jing Zhang; Xianlu Zeng; Ruoxi Wang; Istvan Boldogh; Xueqing Ba

doi:10.1016/j.mcp.2018.04.002

The establishment of methods for free PAR generation and PAR reader detection

Yueshuang Ke, Ke Wang, Hui Xu, Chenxin Wang, Jing Zhang, Xianlu Zeng, Ruoxi Wang, Istvan Boldogh, Xueqing Ba

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

2 Scopus citations

Abstract

Poly (ADP-ribose) polymerase 1 (PARP1) is a DNA damage sensor that catalyzes the poly (ADP-ribose) (PAR) onto a variety of target proteins, such as histones, DSB repair factors and PARP1 itself under consumption of NAD⁺. Besides, PARP1 can affect a variety of proteins in noncovalent modification manner to carry out specific cellular functions. Here, we established a method to generate non-radiolabeled free PAR by PARG moderately cleaving PAR from autoPARylated PARP1, and utilized dot-blot assay to determine the interaction between free PAR and interested proteins. The methods to generate free PAR and detect the noncovalent interactions between proteins and free PAR are nonradioactive and convenient, which will facilitate the studies to explore the significance of PAR reading in various biological processes.

Original language	English (US)
Pages (from-to)	57-60
Number of pages	4
Journal	Molecular and Cellular Probes
Volume	39
DOIs	https://doi.org/10.1016/j.mcp.2018.04.002
State	Published - Jun 2018

Keywords

Dot-blot assay
Free PAR
Noncovalent
PARG
PARP1

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1016/j.mcp.2018.04.002

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title = "The establishment of methods for free PAR generation and PAR reader detection",

abstract = "Poly (ADP-ribose) polymerase 1 (PARP1) is a DNA damage sensor that catalyzes the poly (ADP-ribose) (PAR) onto a variety of target proteins, such as histones, DSB repair factors and PARP1 itself under consumption of NAD+. Besides, PARP1 can affect a variety of proteins in noncovalent modification manner to carry out specific cellular functions. Here, we established a method to generate non-radiolabeled free PAR by PARG moderately cleaving PAR from autoPARylated PARP1, and utilized dot-blot assay to determine the interaction between free PAR and interested proteins. The methods to generate free PAR and detect the noncovalent interactions between proteins and free PAR are nonradioactive and convenient, which will facilitate the studies to explore the significance of PAR reading in various biological processes.",

keywords = "Dot-blot assay, Free PAR, Noncovalent, PARG, PARP1",

author = "Yueshuang Ke and Ke Wang and Hui Xu and Chenxin Wang and Jing Zhang and Xianlu Zeng and Ruoxi Wang and Istvan Boldogh and Xueqing Ba",

note = "Funding Information: This work was supported by grants from the National Natural Science Foundation of China (Grant no. 31371293 and 31571339 to XB), the Natural Science Foundation of Jilin, China (Grant no. 20180101236JC to XB), and grants from United States National Institute of Environmental Health and Sciences (Grant no. RO1 ES018948 to IB), United States National Institute of Allergic and Infectious Diseases (Grant no. AI062885 to IB). Publisher Copyright: {\textcopyright} 2018 Elsevier Ltd",

year = "2018",

month = jun,

doi = "10.1016/j.mcp.2018.04.002",

language = "English (US)",

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T1 - The establishment of methods for free PAR generation and PAR reader detection

AU - Ke, Yueshuang

AU - Wang, Ke

AU - Xu, Hui

AU - Wang, Chenxin

AU - Zhang, Jing

AU - Zeng, Xianlu

AU - Wang, Ruoxi

AU - Boldogh, Istvan

AU - Ba, Xueqing

N1 - Funding Information: This work was supported by grants from the National Natural Science Foundation of China (Grant no. 31371293 and 31571339 to XB), the Natural Science Foundation of Jilin, China (Grant no. 20180101236JC to XB), and grants from United States National Institute of Environmental Health and Sciences (Grant no. RO1 ES018948 to IB), United States National Institute of Allergic and Infectious Diseases (Grant no. AI062885 to IB). Publisher Copyright: © 2018 Elsevier Ltd

PY - 2018/6

Y1 - 2018/6

N2 - Poly (ADP-ribose) polymerase 1 (PARP1) is a DNA damage sensor that catalyzes the poly (ADP-ribose) (PAR) onto a variety of target proteins, such as histones, DSB repair factors and PARP1 itself under consumption of NAD+. Besides, PARP1 can affect a variety of proteins in noncovalent modification manner to carry out specific cellular functions. Here, we established a method to generate non-radiolabeled free PAR by PARG moderately cleaving PAR from autoPARylated PARP1, and utilized dot-blot assay to determine the interaction between free PAR and interested proteins. The methods to generate free PAR and detect the noncovalent interactions between proteins and free PAR are nonradioactive and convenient, which will facilitate the studies to explore the significance of PAR reading in various biological processes.

AB - Poly (ADP-ribose) polymerase 1 (PARP1) is a DNA damage sensor that catalyzes the poly (ADP-ribose) (PAR) onto a variety of target proteins, such as histones, DSB repair factors and PARP1 itself under consumption of NAD+. Besides, PARP1 can affect a variety of proteins in noncovalent modification manner to carry out specific cellular functions. Here, we established a method to generate non-radiolabeled free PAR by PARG moderately cleaving PAR from autoPARylated PARP1, and utilized dot-blot assay to determine the interaction between free PAR and interested proteins. The methods to generate free PAR and detect the noncovalent interactions between proteins and free PAR are nonradioactive and convenient, which will facilitate the studies to explore the significance of PAR reading in various biological processes.

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KW - Noncovalent

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KW - PARP1

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The establishment of methods for free PAR generation and PAR reader detection

Abstract

Keywords

ASJC Scopus subject areas

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