Stable isotope dilution-based accurate comparative quantification of nitrogen-containing metabolites in Arabidopsis thaliana T87 cells using in vivo 15N-isotope enrichment

Jae Kwang Kim, Kazuo Harada, Takeshi Bamba, Ei Ichiro Fukusaki, Akio Kobayashi

研究成果: ジャーナルへの寄稿記事

36 引用 (Scopus)

抄録

Stable isotope dilution-based comparative quantification of nitrogen-containing metabolites for highly sensitive and selective metabolomics was developed using liquid chromatography/mass spectrometry (LC/MS) and 15N-isotope enrichment. We produced metabolically stable isotope-labeled Arabidopsis T87 cells by culturing with 15N-labeled medium. We found that the growth of cells maintained in 15N-labeled medium is very similar to the growth in normal medium, as evidenced by cell morphology, doubling time, and measurement of chlorophyll and carotenoid contents. Complete incorporation of 15N in folate, S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH) in T87 cells was accomplished after culturing for 21 d. Accurate comparative quantification of folate, SAM, and SAH was established by means of LC/MS using the isotopomers of the target metabolites as internal standards. The within- and between-run assay coefficients of variation for the folate, SAM, and SAH levels were all less than 8.5%. Stable isotope labeling by nitrogen source in Arabidopsis T87 cell culture provided simple, inexpensive, and accurate amino acid profiling. This interesting new protocol is valuable for the study of dynamic changes in N-compound pools in cultured cells.

元の言語英語
ページ(範囲)1331-1340
ページ数10
ジャーナルBioscience, Biotechnology and Biochemistry
69
発行部数7
DOI
出版物ステータス出版済み - 9 1 2005

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S-Adenosylhomocysteine
Metabolites
Arabidopsis
Isotopes
S-Adenosylmethionine
Dilution
Nitrogen
Folic Acid
Liquid chromatography
Liquid Chromatography
Mass spectrometry
Mass Spectrometry
Cells
Isotope Labeling
Metabolomics
Carotenoids
Chlorophyll
Growth
Cell culture
Labeling

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

これを引用

Stable isotope dilution-based accurate comparative quantification of nitrogen-containing metabolites in Arabidopsis thaliana T87 cells using in vivo 15N-isotope enrichment. / Kim, Jae Kwang; Harada, Kazuo; Bamba, Takeshi; Fukusaki, Ei Ichiro; Kobayashi, Akio.

:: Bioscience, Biotechnology and Biochemistry, 巻 69, 番号 7, 01.09.2005, p. 1331-1340.

研究成果: ジャーナルへの寄稿記事

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