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Published on: January 22, 2018
Probing Intracellular Yeast Metabolism With Deuterium Magnetic Resonance Spectroscopy
Fatima Anum1, Charbel Assaf1, Farhad Haj Mohamad1
1Section Biomedical Imaging, Molecular Imaging North Competence Center, Department of Radiology and Neuroradiology, University Hospital Schleswig-Holstein, Kiel University, Kiel, Germany.
Deuterium magnetic resonance spectroscopy (DMRS) offers a noninvasive method to track metabolic flux in real-time, overcoming limitations of traditional NMR. This technique successfully monitored nutrient consumption and metabolic changes in yeast cells, demonstrating its versatility.
Area of Science:
- Metabolomics
- Biochemistry
- Spectroscopy
Background:
- Traditional 1H NMR struggles with signal overlap in complex biological samples like cells and tissues.
- Metabolomics aims to capture metabolic states noninvasively, but limitations exist for intact biological systems.
- Nuclear magnetic resonance (NMR) is a key noninvasive technique in metabolomics.
Purpose of the Study:
- To introduce and validate Deuterium Magnetic Resonance Spectroscopy (DMRS) as a method for metabolic flux analysis.
- To demonstrate DMRS's capability in tracking real-time metabolite consumption and metabolic pathway alterations.
- To assess DMRS parameters for optimizing measurements in biological systems.
Main Methods:
- Utilized deuterium-labeled tracers to circumvent background interference in NMR spectroscopy.
- Measured key spectroscopic parameters (T1, T2, chemical shifts) for 26 deuterated compounds.
- Applied DMRS to Saccharomyces cerevisiae (baker's yeast) under varied substrate concentrations and osmotic pressures.
Main Results:
- Successfully tracked the real-time consumption of pyruvate, glucose, fumarate, acetone, and nicotinamide in yeast.
- Demonstrated the ability to redirect yeast metabolism by manipulating pyruvate levels and buffer density.
- Established optimal DMRS settings through characterization of deuterated compounds.
Conclusions:
- DMRS is a robust and versatile tool for dissecting metabolic transformations noninvasively.
- The study highlights DMRS's potential for real-time metabolic flux analysis in accessible model systems like yeast.
- DMRS enables efficient metabolic flux analysis without requiring specialized equipment.
