Phosphorus NMR and Its Application to Metabolomics

Fatema Bhinderwala, Paula Evans, Kaleb Jones, Benjamin R. Laws, Thomas G. Smith, Martha Morton, Robert Powers

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Stable isotopes are routinely employed by NMR metabolomics to highlight specific metabolic processes and to monitor pathway flux. 13C-carbon and 15N-nitrogen labeled nutrients are convenient sources of isotope tracers and are commonly added as supplements to a variety of biological systems ranging from cell cultures to animal models. Unlike 13C and 15N, 31P-phosphorus is a naturally abundant and NMR active isotope that does not require an external supplemental source. To date, 31P NMR has seen limited usage in metabolomics because of a lack of reference spectra, difficulties in sample preparation, and an absence of two-dimensional (2D) NMR experiments, but 31P NMR has the potential of expanding the coverage of the metabolome by detecting phosphorus-containing metabolites. Phosphorylated metabolites regulate key cellular processes, serve as a surrogate for intracellular pH conditions, and provide a measure of a cell's metabolic energy and redox state, among other processes. Thus, incorporating 31P NMR into a metabolomics investigation will enable the detection of these key cellular processes. To facilitate the application of 31P NMR in metabolomics, we present a unified protocol that allows for the simultaneous and efficient detection of 1H-, 13C-, 15N-, and 31P-labeled metabolites. The protocol includes the application of a 2D 1H-31P HSQC-TOCSY experiment to detect 31P-labeled metabolites from heterogeneous biological mixtures, methods for sample preparation to detect 1H-, 13C-, 15N-, and 31P-labeled metabolites from a single NMR sample, and a data set of one-dimensional (1D) 31P NMR and 2D 1H-31P HSQC-TOCSY spectra of 38 common phosphorus-containing metabolites to assist in metabolite assignments.

Original languageEnglish (US)
Pages (from-to)9536-9545
Number of pages10
JournalAnalytical chemistry
Volume92
Issue number14
DOIs
StatePublished - Jul 21 2020

ASJC Scopus subject areas

  • Analytical Chemistry

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    Bhinderwala, F., Evans, P., Jones, K., Laws, B. R., Smith, T. G., Morton, M., & Powers, R. (2020). Phosphorus NMR and Its Application to Metabolomics. Analytical chemistry, 92(14), 9536-9545. https://doi.org/10.1021/acs.analchem.0c00591