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 language | English (US) |
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Pages (from-to) | 24-28 |
Number of pages | 5 |
Journal | Analytical Biochemistry |
Volume | 282 |
Issue number | 1 |
DOIs | |
State | Published - Jun 15 2000 |
Externally published | Yes |
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology