論文

査読有り
2008年11月

Systematic NMR Analysis of Stable Isotope Labeled Metabolite Mixtures in Plant and Animal Systems: Coarse Grained Views of Metabolic Pathways

PLOS ONE
  • Eisuke Chikayama
  • ,
  • Michitaka Suto
  • ,
  • Takashi Nishihara
  • ,
  • Kazuo Shinozaki
  • ,
  • Takashi Hirayama
  • ,
  • Jun Kikuchi

3
11
開始ページ
e3805
終了ページ
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1371/journal.pone.0003805
出版者・発行元
PUBLIC LIBRARY SCIENCE

Background: Metabolic phenotyping has become an important 'bird's-eye-view' technology which can be applied to higher organisms, such as model plant and animal systems in the post-genomics and proteomics era. Although genotyping technology has expanded greatly over the past decade, metabolic phenotyping has languished due to the difficulty of 'top-down' chemical analyses. Here, we describe a systematic NMR methodology for stable isotope-labeling and analysis of metabolite mixtures in plant and animal systems.
Methodology/Principal Findings: The analysis method includes a stable isotope labeling technique for use in living organisms; a systematic method for simultaneously identifying a large number of metabolites by using a newly developed HSQC-based metabolite chemical shift database combined with heteronuclear multidimensional NMR spectroscopy; Principal Components Analysis; and a visualization method using a coarse-grained overview of the metabolic system. The database contains more than 1000 (1)H and (13)C chemical shifts corresponding to 142 metabolites measured under identical physicochemical conditions. Using the stable isotope labeling technique in Arabidopsis T87 cultured cells and Bombyx mori, we systematically detected >450 HSQC peaks in each (13)C-HSQC spectrum derived from model plant, Arabidopsis T87 cultured cells and the invertebrate animal model Bombyx mori. Furthermore, for the first time, efficient (13)C labeling has allowed reliable signal assignment using analytical separation techniques such as 3D HCCH-COSY spectra in higher organism extracts.
Conclusions/Significance: Overall physiological changes could be detected and categorized in relation to a critical developmental phase change in B. mori by coarse-grained representations in which the organization of metabolic pathways related to a specific developmental phase was visualized on the basis of constituent changes of 56 identified metabolites. Based on the observed intensities of (13)C atoms of given metabolites on development-dependent changes in the 56 identified (13)C-HSQC signals, we have determined the changes in metabolic networks that are associated with energy and nitrogen metabolism.

リンク情報
DOI
https://doi.org/10.1371/journal.pone.0003805
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/19030231
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000265451100005&DestApp=WOS_CPL
URL
http://orcid.org/0000-0002-6809-394X
ID情報
  • DOI : 10.1371/journal.pone.0003805
  • ISSN : 1932-6203
  • ORCIDのPut Code : 23626361
  • PubMed ID : 19030231
  • Web of Science ID : WOS:000265451100005
  • ORCIDで取得されたその他外部ID : a:1:{i:0;a:1:{s:8:"other-id";s:19:"WOS:000265451100005";}}

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