Binding kinetics and activity of human poly (ADP-ribose) polymerase-1 on oligo-deoxyribonucleotide substrates

Timothy J. Jorgensen, Kevin Chen, Sergey Chasovskikh, Rabindra Roy, Anatoly Dritschilo, Aykut Üren

Research output: Contribution to journalArticlepeer-review

16 Scopus citations


Poly(ADP-ribose) polymerase-1 (PARP-1) is a mammalian enzyme that attaches long branching chains of ADP-ribose to specific nuclear proteins, including itself. Because its activity in vitro is dependent upon interaction with broken DNA, it has been postulated that PARP-1 plays an important role in DNA strand-break repair in vivo. The exact mechanism of binding to DNA and the structural determinants of binding remain to be defined, but regions of transition from single-stranded to double-strandedness may be important recognition sites. Here we employ surface plasmon resonance (SPR) to investigate this hypothesis. Oligodeoxynucleotide (ODN) substrates that mimic DNA with different degrees of single-strandedness were used for measurements of both PARP-1/DNA binding kinetics and PARP-1's enzyme activities. We found that binding correlated with activity, but was unrelated to single-strandedness of the ODN. Instead, PARP-1 binding and activity were highest on ODNs that modeled a DNA double-strand break (DSB). These results provide support for PARP-1 recognizing and binding DSBs in a manner that is independent of single-stranded features, and demonstrate the usefulness of SPR for simultaneously investigating both PARP-1 binding and PARP-1 auto-poly(ADP-ribosyl)ation activities within the same in vitro system.

Original languageEnglish (US)
Pages (from-to)446-452
Number of pages7
JournalJournal of Molecular Recognition
Issue number6
StatePublished - Nov 2009
Externally publishedYes


  • Binding kinetics
  • DNA double-strand breaks
  • DNA repair
  • Non-homologous end joining
  • PARP-1
  • Poly(ADP-ribose) polymerase-1
  • Poly(ADP-ribosyl)ation
  • Surface plasmon resonance

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology


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