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Small-Molecule Screening Assay for Mono-ADP-Ribosyltransferases

  • Teemu Haikarainen
  • Sudarshan Murthy
  • Mirko M. Maksimainen
  • Lari Lehtiö
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1813)

Abstract

Mono-ADP-ribosyltransferases of the PARP/ARTD enzyme family are enzymes catalyzing the transfer of a single ADP-ribose unit to target proteins. The enzymes have various roles in vital cellular processes such as DNA repair and transcription, and many of the enzymes are linked to cancer-relevant functions. Thus inhibition of the enzymes is a potential way to discover and develop new drugs against cancer. Here we describe an activity-based screening assay for mono-ADP-ribosyltransferases. The assay utilizes the natural substrate of the enzymes, NAD+, and it is based on chemically converting the leftover substrate to a fluorophore and measuring its relative concentration after the enzymatic reaction. The assay is homogenous, robust, and cost-effective and, most importantly, applicable to mono-ADP-ribosyltransferases as well as poly-ADP-ribosyltransferases for screening of small-molecule inhibitors against the enzymes.

Key words

PARP ARTD Mono-ADP-ribosyltransferase Screening assay Inhibitor 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Teemu Haikarainen
    • 1
  • Sudarshan Murthy
    • 1
  • Mirko M. Maksimainen
    • 1
  • Lari Lehtiö
    • 1
  1. 1.Faculty of Biochemistry and Molecular Medicine, Biocenter OuluUniversity of OuluOuluFinland

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