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

doi:10.1021/bc015560h

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 journal › Article › peer-review

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 language	English
Pages (from-to)	163-166
Number of pages	4
Journal	Bioconjugate Chemistry
Volume	13
Issue number	2
DOIs	https://doi.org/10.1021/bc015560h
Publication status	Published - Apr 13 2002
Externally published	Yes

ASJC Scopus subject areas

Biotechnology
Bioengineering
Biomedical Engineering
Pharmacology
Pharmaceutical Science
Organic Chemistry

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10.1021/bc015560h

Cite this

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

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.",

author = "Takenori Tomohiro and Sawada, {Jun Ichi} and Chika Sawa and Hironori Nakura and Shuhei Yoshida and Masato Kodaka and Mamoru Hatakeyama and Haruma Kawaguchi and Hiroshi Handa and Hiroaki Okuno",

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AU - Tomohiro, Takenori

AU - Sawada, Jun Ichi

AU - Sawa, Chika

AU - Nakura, Hironori

AU - Yoshida, Shuhei

AU - Kodaka, Masato

AU - Hatakeyama, Mamoru

AU - Kawaguchi, Haruma

AU - Handa, Hiroshi

AU - Okuno, Hiroaki

PY - 2002/4/13

Y1 - 2002/4/13

N2 - 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.

AB - 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.

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

Abstract

ASJC Scopus subject areas

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