NMR-based metabolite studies with 15N amino acids
| Autor: | Arnon Lavie, Michael Caffrey, Benjamin E. Ramirez, Matthew A. Durst |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Magnetic Resonance Spectroscopy Metabolite lcsh:Medicine Article Enzyme catalysis 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine NMR spectroscopy Cell Line Tumor Ammonium Compounds Humans Amino Acids lcsh:Science chemistry.chemical_classification Multidisciplinary Nitrogen Isotopes Biological techniques lcsh:R Temperature Metabolism Amino acid 030104 developmental biology chemistry Biochemistry Cell culture Nucleic acid lcsh:Q 030217 neurology & neurosurgery |
| Zdroj: | Scientific Reports, Vol 9, Iss 1, Pp 1-5 (2019) Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-019-49208-8 |
| Popis: | 15N labeled amino acids are routinely used to label proteins or nucleic acids for study by NMR. However, NMR studies of 15N labeled amino acids in metabolite studies have not been pursued extensively, presumably due to line broadening present under standard experimental conditions. In this work, we show that lowering the temperature to −5 °C allows facile characterization of 15N-labeled amino acids. Further, we show that this technique can be exploited to measure 15NH3 produced in an enzyme catalyzed reaction and the transport and metabolism of individual amino acids in mammalian cell culture. With respect to 13C-labeled amino acids, 15N-labeled amino acids are less costly and enable direct characterization of nitrogen metabolism in complex biological systems by NMR. In summary, the present work significantly expands the metabolite pools and their reactions for study by NMR. |
| Databáze: | OpenAIRE |
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