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

• YAMAZAKI Kenichi

  Laboratory of Bio-analytical Chemistry, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Josai University

• IKEGUCHI Yoshihiko

  Laboratory of Bio-analytical Chemistry, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Josai University

• 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.

収録刊行物

• Analytical Sciences

  Analytical Sciences 28 (6), 621-624, 2012

  社団法人 日本分析化学会

キーワード

• Aminopropyltrnasferase
• spermine synthase
• spermidine synthase
• Etheno-derivatization
• methylthioadenosine

詳細情報

• CRID: 1390282679236452480
• NII論文ID: 10030758186
• NII書誌ID: AA10500785
• DOI: 10.2116/analsci.28.621
• COI: 1:STN:280:DC%2BC38jksVKisA%3D%3D
• ISSN: 13482246
• ISSN: 09106340
• NDL書誌ID: 023689241
• PubMed: 22729051
• Web Site: http://libir.josai.ac.jp/il/meta_pub/G0000284repository_JOS-analsci.28.621
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I023689241
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.28.621.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.28.621/fulltext.html
• Web Site: https://www.jstage.jst.go.jp/article/analsci/28/6/28_621/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/2013046774
• 本文言語コード: en
• データソース種別:
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  • IRDB
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
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