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 journalArticle

    1 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 languageEnglish (US)
    JournalMolecular and Cellular Probes
    DOIs
    StateAccepted/In press - Jan 1 2018

    Keywords

    • Dot-blot assay
    • Free PAR
    • Noncovalent
    • PARG
    • PARP1

    ASJC Scopus subject areas

    • Molecular Biology
    • Cell Biology

    Fingerprint Dive into the research topics of 'The establishment of methods for free PAR generation and PAR reader detection'. Together they form a unique fingerprint.

  • Cite this