Phosphorus NMR and Its Application to Metabolomics
- 12 June 2020
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 92 (14), 9536-9545
- https://doi.org/10.1021/acs.analchem.0c00591
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.Keywords
Funding Information
- Division of Biological Infrastructure (1660921)
- National Institute of General Medical Sciences (P20 GM113126, P30 GM103335)
This publication has 35 references indexed in Scilit:
- HMDB 3.0—The Human Metabolome Database in 2013Nucleic Acids Research, 2012
- TOCCATA: A Customized Carbon Total Correlation Spectroscopy NMR Metabolomics DatabaseAnalytical Chemistry, 2012
- Predicting the in Vivo Mechanism of Action for Drug Leads Using NMR MetabolomicsACS Chemical Biology, 2011
- Highly routinely reproducible alignment of 1H NMR spectral peaks of metabolites in huge sets of urinesAnalytica Chimica Acta, 2011
- DNA structures from phosphate chemical shiftsNucleic Acids Research, 2009
- BioMagResBank (BMRB) as a partner in the Worldwide Protein Data Bank (wwPDB): new policies affecting biomolecular NMR depositionsJournal of Biomolecular NMR, 2008
- Three-Dimensional Triple-Resonance 1H, 13C, 31P Experiment: Sequential Through-Bond Correlation of Ribose Protons and Intervening Phosphorus along the RNA Oligonucleotide BackboneJournal of the American Chemical Society, 1994
- Phosphorus-31 nuclear magnetic resonance of dihydroxyacetone phosphate in the presence of triosephosphate isomerase. The question of nonproductive binding of the substrate hydrateBiochemistry, 1977
- 31P chemical shifts in phosphate diester monoanions. Bond angle and torsional angle effectsBiochemical and Biophysical Research Communications, 1975
- Kinetics of adenosine-5′-triphosphate hydrolysisJournal of Inorganic and Nuclear Chemistry, 1974