Purification and characterization of poly(ADP-ribose) polymerase from Sarcophaga peregrina

Miyoko Ikejima, Tadashige Nozaki, Shoichiro Kurata, Shunji Natori, Hiroyasu Esumi, Takashi Sugimura, Mitsuko Masutani

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1 Citation (Scopus)


Poly(ADP-ribose) polymerase (PARP) was purified from embryonic NIH-Sape-4 cells of Sarcophaga peregrina almost to homogeneity. The purification was carried out using phosphocellulose and 3-aminobenzamide-Sepharose column chromatographies. The purified enzyme had an apparent molecular weight of 113 kDa on SDS-polyacrylamide gel electrophoresis. The addition of DNA was essential for the activity of the purified enzyme, and the reaction was stimulated by the presence of histone and magnesium ion. The Km for NAD was 43 μM and the optimum temperature for the PARP activity ranged from 20-25 °C. MALDI (matrix-associated laser desorption ionisation)-TOF (time-of-flight) mass spectrometric analysis gave a signal at 112,402. This value was close to 113,033, the predicted mass from its cDNA sequence. This suggests that full length PARP is present as a major native form without extensive post-translational modification.

Original languageEnglish
Pages (from-to)282-285
Number of pages4
JournalProceedings of the Japan Academy Series B: Physical and Biological Sciences
Issue number9
Publication statusPublished - 2002 Nov


  • Mass spectrometry
  • Poly(ADP-ribose) polymerase
  • Purification
  • Sarcophaga

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Physics and Astronomy(all)


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