Total analysis and purification of cellular proteins binding to cisplatin-damaged DNA using submicron beads

Takenori Tomohiro, Jun Ichi Sawada, Chika Sawa, Hironori Nakura, Shuhei Yoshida, Masato Kodaka, Mamoru Hatakeyama, Haruma Kawaguchi, Hiroshi Handa, Hiroaki Okuno

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

A high-performance affinity purification technique has been developed for cisplatin (CDDP)-damaged DNA binding proteins directly from crude nuclear extracts of HeLaS3 cell using novel submicron beads synthesized by copolymerization of styrene and glycidyl methacrylate (GMA). The beads dramatically decreased both nonspecific protein adsorption on solid surfaces and elution volume and simplified the handling procedure. Preparation of the beads for purification was carried out by immobilization of telomeric repeats, (TTAGGG)n, on the surface after the reaction with CDDP. At least nine proteins clearly showed higher affinity to CDDP-DNA and were identified by amino acid sequence analysis including HMGB (high mobility group), hUBF (human upstream binding factor), and Ku autoantigen, which were previously reported to be components of CDDP-damaged DNA binding proteins.

Original languageEnglish
Pages (from-to)163-166
Number of pages4
JournalBioconjugate Chemistry
Volume13
Issue number2
DOIs
Publication statusPublished - Apr 13 2002
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

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  • Cite this

    Tomohiro, T., Sawada, J. I., Sawa, C., Nakura, H., Yoshida, S., Kodaka, M., Hatakeyama, M., Kawaguchi, H., Handa, H., & Okuno, H. (2002). Total analysis and purification of cellular proteins binding to cisplatin-damaged DNA using submicron beads. Bioconjugate Chemistry, 13(2), 163-166. https://doi.org/10.1021/bc015560h