Determination of cellular aminopropyltransferase activity using precolumn fluorescent etheno-derivatization with high-performance liquid chromatography

Kenichi Yamazaki, Yoshihiko Ikeguchi, Takuya Niwa, Kaoru Hayashi, Takahiro Iwaki, Ikumi Ishii, Masaru Niitsu, Anthony Pegg, Akira Shirahata

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)621-624
Number of pages4
JournalAnalytical Sciences
Volume28
Issue number6
DOIs
StatePublished - Jan 1 2012

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Spermidine Synthase
High performance liquid chromatography
Spermine Synthase
Spermidine
Spermine
Polyamines
Cell growth
Oleic Acid
Calibration
Lipids
Liquids

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

Cite this

Yamazaki, Kenichi ; Ikeguchi, Yoshihiko ; Niwa, Takuya ; Hayashi, Kaoru ; Iwaki, Takahiro ; Ishii, Ikumi ; Niitsu, Masaru ; Pegg, Anthony ; Shirahata, Akira. / Determination of cellular aminopropyltransferase activity using precolumn fluorescent etheno-derivatization with high-performance liquid chromatography. In: Analytical Sciences. 2012 ; Vol. 28, No. 6. pp. 621-624.
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Determination of cellular aminopropyltransferase activity using precolumn fluorescent etheno-derivatization with high-performance liquid chromatography. / Yamazaki, Kenichi; Ikeguchi, Yoshihiko; Niwa, Takuya; Hayashi, Kaoru; Iwaki, Takahiro; Ishii, Ikumi; Niitsu, Masaru; Pegg, Anthony; Shirahata, Akira.

In: Analytical Sciences, Vol. 28, No. 6, 01.01.2012, p. 621-624.

Research output: Contribution to journalArticle

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AU - Iwaki, Takahiro

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AU - Niitsu, Masaru

AU - Pegg, Anthony

AU - Shirahata, Akira

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