Expanding the Coverage of the Metabolome with Nitrogen-Based NMR

Fatema Bhinderwala, Samantha Lonergan, Jade Woods, Chunyi Zhou, Paul D. Fey, Robert Powers

Research output: Contribution to journalArticle

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Abstract

Isotopically labeling a metabolite and tracing its metabolic fate has provided invaluable insights about the role of metabolism in human diseases in addition to a variety of other issues. 13C-labeled metabolite tracers or unlabeled 1H-based NMR experiments are currently the most common application of NMR to metabolomics studies. Unfortunately, the coverage of the metabolome has been consequently limited to the most abundant carbon-containing metabolites. To expand the coverage of the metabolome and enhance the impact of metabolomics studies, we present a protocol for 15N-labeled metabolite tracer experiments that may also be combined with routine 13C tracer experiments to simultaneously detect both 15N- and 13C-labeled metabolites in metabolic samples. A database consisting of 2D 1H-15N HSQC natural-abundance spectra of 50 nitrogen-containing metabolites are also presented to facilitate the assignment of 15N-labeled metabolites. The methodology is demonstrated by labeling Escherichia coli and Staphylococcus aureus metabolomes with 15N1-ammonium chloride, 15N4-arginine, and 13C2-acetate. Efficient 15N and 13C metabolite labeling and identification were achieved utilizing standard cell culture and sample preparation protocols.

Original languageEnglish (US)
Pages (from-to)4521-4528
Number of pages8
JournalAnalytical chemistry
Volume90
Issue number7
DOIs
StatePublished - Apr 3 2018

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ASJC Scopus subject areas

  • Analytical Chemistry

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