New method for the quantitative determination of major protein carbonyls, α-aminoadipic and γ-glutamic semialdehydes: Investigation of the formation mechanism and chemical nature in vitro and in vivo

Mitsugu Akagawa, Daisuke Sasaki, Yoshihisa Ishii, Yayoi Kurota, Mari Yotsu-Yamashita, Koji Uchida, Kyozo Suyama

    研究成果: Article査読

    51 被引用数 (Scopus)

    抄録

    α-Aminoadipic semialdehyde (AAS) and γ-glutamic semialdehyde (GGS) are identified as the major carbonyl products in oxidized proteins. To elucidate the formation pathway of AAS and GGS in vivo, we developed and validated a new quantification method. AAS and GGS in proteins were derivatized by reductive amination with NaCNBH3 and p-aminobenzoic acid, a fluorescent reagent, followed by acid hydrolysis. It is noteworthy that the fluorescent derivatives were completely stable during acid hydrolysis. The present method permitted the specific, accurate, and sensitive quantification of both semialdehydes by fluorometric high-performance liquid chromatography. Analysis of proteins oxidized by various oxidation systems revealed that AAS and GGS are notably generated by the reaction of proteins with .OH, which is produced by metal-catalyzed oxidation (MCO). Furthermore, exposure of transferrin and human plasma to ascorbic acid and H2O2 significantly promoted the formation of AAS and GGS in vitro, suggesting that both semialdehydes can be generated by MCO in vivo. We also demonstrated their generation through oxidative stress induced by acute iron overload in vivo. In this paper, we describe this analytical technique for simple and precise measurement of AAS and GGS and discuss their formation mechanism in vivo.

    本文言語English
    ページ(範囲)1059-1065
    ページ数7
    ジャーナルChemical Research in Toxicology
    19
    8
    DOI
    出版ステータスPublished - 2006 8月

    ASJC Scopus subject areas

    • 毒物学

    フィンガープリント

    「New method for the quantitative determination of major protein carbonyls, α-aminoadipic and γ-glutamic semialdehydes: Investigation of the formation mechanism and chemical nature in vitro and in vivo」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

    引用スタイル