Determination of Cellular Aminopropyltransferase Activity Using Precolumn Fluorescent Etheno-Derivatization with High-Performance Liquid Chromatography

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著者

    • NIWA Takuya [他] NIWA Takuya
    • Laboratory of Bio-analytical Chemistry, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Josai University
    • HAYASHI Kaoru
    • Laboratory of Bio-analytical Chemistry, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Josai University
    • IWAKI Takahiro
    • Laboratory of Bio-analytical Chemistry, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Josai University
    • ISHII Ikumi
    • Laboratory of Bio-analytical Chemistry, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Josai University
    • NIITSU Masaru
    • Laboratory of Bio-analytical Chemistry, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Josai University
    • PEGG Anthony E.
    • Department of Cellular and Molecular Physiology, Milton S. Hershey Medical Center, Pennsylvania State University College of Medicine
    • SHIRAHATA Akira
    • Laboratory of Bio-analytical Chemistry, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Josai University

抄録

Polyamines such as spermidine (Spd) and spermine (Spm), produced by aminopropyltransferase (Apt), play roles in cell growth and differentiation. A sensitive and simple fluorometric high-performance liquid chromatographic determination for Apt activity of spermidine synthase (Spdsyn) and spermine synthase (Spmsyn) was developed in order to examine cellular functions of polyamine synthesis. The derivatization procedure for methylthioadenosine (MTA) produced from decarboxylated S-adenosylmethionine by Apt was the reaction with 2-chloroacetaldehyde to give fluorescent 1, N6-etheno methylthioadenosine. The reaction conditions for derivatization were optimized. A calibration curve was established, ranging from 0.01 to 25 pmol. Quantification of derivatized MTA was confirmed to be identical to Spd or Spm production. The developed method determined Spdsyn and Spmsyn activities in HepG2 cells treated with oleic acid as a cellular lipid accumulation model.

Polyamines such as spermidine (Spd) and spermine (Spm), produced by aminopropyltransferase (Apt), play roles in cell growth and differentiation. A sensitive and simple fluorometric high-performance liquid chromatographic determination for Apt activity of spermidine synthase (Spdsyn) and spermine synthase (Spmsyn) was developed in order to examine cellular functions of polyamine synthesis. The derivatization procedure for methylthioadenosine (MTA) produced from decarboxylated <i>S</i>-adenosylmethionine by Apt was the reaction with 2-chloroacetaldehyde to give fluorescent 1, <i>N</i><sup>6</sup>-etheno methylthioadenosine. The reaction conditions for derivatization were optimized. A calibration curve was established, ranging from 0.01 to 25 pmol. Quantification of derivatized MTA was confirmed to be identical to Spd or Spm production. The developed method determined Spdsyn and Spmsyn activities in HepG2 cells treated with oleic acid as a cellular lipid accumulation model.

収録刊行物

  • Analytical sciences : the international journal of the Japan Society for Analytical Chemistry

    Analytical sciences : the international journal of the Japan Society for Analytical Chemistry 28(6), 621-624, 2012-06-10

    日本分析化学会

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各種コード

  • NII論文ID(NAID)
    10030758186
  • NII書誌ID(NCID)
    AA10500785
  • 本文言語コード
    ENG
  • 資料種別
    NOT
  • ISSN
    09106340
  • NDL 記事登録ID
    023689241
  • NDL 請求記号
    Z54-F482
  • データ提供元
    CJP書誌  NDL  IR  J-STAGE 
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