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Towards dynamic metabolic network measurements by multi-dimensional NMR-based fluxomics by Yasuyo Sekiyama; Jun Kikuchi is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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Novel technologies for measuring biological systems and methods for visualizing data have led to a revolution in the life sciences. Nuclear magnetic resonance (NMR) techniques can provide information on metabolite structure and metabolic dynamics at the atomic level. We have been developing a new method for measuring the dynamic metabolic network of crude extracts that combines [(13)C(6)]glucose stable isotope labeling of Arabidopsis thaliana and multi-dimensional heteronuclear NMR analysis, whereas most conventional metabolic flux analyses examine proteinogenic amino acids that are specifically labeled with partially labeled substrates such as [2-(13)C(1)]glucose or 10% [(13)C(6)]glucose. To show the validity of our method, we investigated how to obtain information about biochemical reactions, C-C bond formation, and the cleavage of the main metabolites, such as free amino acids, in crude extracts based on the analysis of the (13)C-(13)C coupling pattern in 2D-NMR spectra. For example, the combination of different extraction solvents allows one to distinguish complicated (13)C-(13)C fine couplings at the C2 position of amino acids. As another approach, f1-f3 projection of the HCAC

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Author

Yasuyo Sekiyama; Jun Kikuchi

Publisher

Elsevier Science; Elsevier ; Elsevier BV (ISSN 0031-9422)

Published

2007

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)