• Medientyp: E-Artikel
  • Titel: Isotopolog Perturbation Techniques for Metabolic Networks: Metabolic Recycling of Nutritional Glucose in Drosophila melanogaster
  • Beteiligte: Eisenreich, Wolfgang; Ettenhuber, Christian; Laupitz, Ralf; Theus, Cornelia; Bacher, Adelbert; Arigoni, Duilio
  • Erschienen: National Academy of Sciences, 2004
  • Erschienen in: Proceedings of the National Academy of Sciences of the United States of America
  • Sprache: Englisch
  • ISSN: 0027-8424
  • Schlagwörter: Biological Sciences
  • Entstehung:
  • Anmerkungen:
  • Beschreibung: <p>Drosophila melanogaster strain Oregon-R* was grown on standard medium supplemented with [ U-&lt;sup&gt;13&lt;/sup&gt; C&lt;sub&gt;6&lt;/sub&gt;]glucose. One to two days after hatching, flies were extracted with water. Glucose was isolated chromatographically from the extract and was analyzed by &lt;sup&gt;13&lt;/sup&gt; C NMR spectroscopy. All &lt;sup&gt;13&lt;/sup&gt; C signals of the isolated glucose were multiplets arising by &lt;sup&gt;13&lt;/sup&gt; C&lt;sup&gt;13&lt;/sup&gt; C coupling. Based on a comprehensive analysis of the coupling constants and heavy isotope shifts in glucose, the integrals of individual &lt;sup&gt;13&lt;/sup&gt; C signal patterns afforded the concentrations of certain groups of &lt;sup&gt;13&lt;/sup&gt; C isotopologs. These data were deconvoluted by a genetic algorithm affording the abundances of all single-labeled and of 15 multiply labeled isotopologs. Among the latter group, seven isotopologs were found at concentrations &gt;0.1 mol % with [1,2-&lt;sup&gt;13&lt;/sup&gt; C&lt;sub&gt;2&lt;/sub&gt;]glucose as the most prominent species. The multiply &lt;sup&gt;13&lt;/sup&gt; C-labeled glucose isotopologs are caused by metabolic remodeling of the proffered glucose via a complex network of catabolic and anabolic processes involving glycolysis and/or passage through the pentose phosphate, the Cori cycle and/or the citrate cycle. The perturbation method described can be adapted to a wide variety of experimental systems and isotope-labeled precursors.</p>
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