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Continuous in vivo metabolism by NMR

ABSTRACT: Metabolomics relies on analytical methods to provide holistic information about metabolites, their distributions across samples, and their underlying dynamic properties. The latter is gaining increasing attention due to advances in modeling and new analytical methods that provide dense time-series data. We extended high-resolution-magic angle spinning (HR-MAS) NMR—an established technique to measure metabolites from tissues and live organisms—into a flexible, untargeted, and continuous recording of in vivo metabolism. We call this technique “continuous in vivo metabolism by NMR” (CIVM-NMR). We used isotope-edited CIVM-NMR to reproduce a recent amino acid flux result in chronic lymphoid leukemia cells. We then collected untargeted CIVM-NMR datasets for Neurospora crassa, a classic multicellular model of biochemistry, genetics, and metabolism. CIVM-NMR requires virtually no sample preparation and allows for continuous collection of data over hours to days at ~4-min temporal resolution with little noise. CIVM-NMR provided real-time measurements that unambiguously reproduced the direction of flux of branched-chain amino acid accumulation in leukemia cells. It also revealed the dynamics of central carbon metabolism, amino acid metabolism, energy storage molecules, and lipid and cell wall precursors in N. crassa. CIVM-NMR is simple and readily adapted to different types of cells and microorganisms, making it ideally suited to experimentally complement kinetic models of metabolism for diverse biological systems.

ORGANISM(S): Fungi Neurospora Crassa

TISSUE(S): Mycelia In Media

SUBMITTER: Michael Judge

PROVIDER: ST001103 | MetabolomicsWorkbench | Tue Nov 27 00:00:00 GMT 2018

REPOSITORIES: MetabolomicsWorkbench

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