A new type of O2--generating tool for oxidative stress studies by remodeling neutrophil NADPH oxidase

Minoru Tamura, Teruaki Nagasawa, Tomoyuki Tange, Kei Miyano, Shin Ichi Kobayashi, Toshiki Nakai, Satoshi Miura

研究成果: Contribution to journalArticle査読

3 被引用数 (Scopus)

抄録

The effects of reactive oxygen species on cells have attracted much attention in relation to redox regulation and oxidative stress-related diseases. Superoxide (O2-) is the reactive oxygen species primarily formed in biological systems. However, no convenient O2 --generating device has been available for use in cell or tissue culture. The neutrophil NADPH oxidase, a professional enzyme for killing bacteria, has a high ability to produce O2-. However, the cell-free activation process requires several protein factors and an anionic amphiphile, and moreover, the activation is transient. To utilize the enzyme as an O2- generator, we improved the cell-free activation method by remodeling regulatory components, optimizing lipid composition, and modifying the mixing conditions. We established a new method to produce an active enzyme that is stable, efficient, and preservable. As an application, we examined the effect of the device on cultured HEK293 cells and observed that it caused cell death. This system has several advantages over the xanthine oxidase system often used. The new device will be useful for studies of oxidative stress and related diseases.

本文言語英語
ページ(範囲)421-429
ページ数9
ジャーナルJournal of Biotechnology
120
4
DOI
出版ステータス出版済み - 12 6 2005

All Science Journal Classification (ASJC) codes

  • バイオテクノロジー
  • バイオエンジニアリング
  • 応用微生物学とバイオテクノロジー

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