Detection of Metabolic Fluxes of O and H Atoms into Intracellular Water in Mammalian Cells
Metabolic processes result in the release and exchange of H and O atoms from organic material as well as some inorganic salts and gases. These fluxes of H and O atoms into intracellular water result in an isotopic gradient that can be measured experimentally. Using isotope ratio mass spectroscopy, we revealed that slightly over 50% of the H and O atoms in the intracellular water of exponentially-growing cultured Rat-1 fibroblasts were isotopically distinct from growth medium water. We then employed infrared spectromicroscopy to detect in real time the flux of H atoms in these same cells. Importantly, both of these techniques indicate that the H and O fluxes are dependent on metabolic processes; cells that are in lag phase or are quiescent exhibit a much smaller flux. In addition, water extracted from the muscle tissue of rats contained a population of H and O atoms that were isotopically distinct from body, consistent with the results obtained using the cultured Rat-1 fibroblasts. Together these data demonstrate that metabolic processes produce fluxes of H and O atoms into intracellular water, and that these fluxes can be detected and measured in both cultured mammalian cells and in mammalian tissue.
- Research Organization:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 1053347
- Report Number(s):
- PNNL-SA-84966; KP1601010
- Journal Information:
- Journal of Biological Chemistry, 7(7):Article No. e39685, Journal Name: Journal of Biological Chemistry, 7(7):Article No. e39685
- Country of Publication:
- United States
- Language:
- English
Similar Records
Metabolic Processes Account for the Majority of Intracellular Water in Log-Phase Escherichia coli Cells as Revealed by Hydrogen Isotopes
Degradation of horseradish peroxidase after microinjection into mammalian cells