A scintillation proximity assay for poly(ADP-ribose) polymerase

Anissa Cheung, Jie Zhang

Research output: Contribution to journalArticlepeer-review

Abstract

Poly(ADP-ribose) polymerase (PARP) is an abundant nuclear protein in most of the eukaryotic tissues. When activated by DNA damage, PARP synthesizes poly(ADP-ribose) from NAD. Conventional radioactive PARP enzyme assay requires the separation of the polymer product from the NAD substrate, a rate-limiting step that hampers large-scale chemical library screening to identify novel small-molecule PARP inhibitors. By using biotinylated NAD, we have developed a scintillation proximity assay (SPA) for PARP. We demonstrated that PARP can incorporate the biotinylated ADP-ribose units into the radioactive poly(ADP-ribose) polymer, which can directly bind and excite the streptavidin-conjugated scintillation beads. PARP-SPA can be readily adapted to a 96-well format for automatic high-throughput screening for PARP inhibitors. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)24-28
Number of pages5
JournalAnalytical Biochemistry
Volume282
Issue number1
DOIs
StatePublished - Jun 15 2000
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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