Our research into cell metabolism is underpinned by the Metabolic Tracer Analysis Core (MTAC), which specialises in high resolution methodologies to investigate metabolic pathway activity in health and disease. Central to our approach is the use of stable isotope-enriched nutrients to examine their use under different conditions – for example, 13C6-glucose or 13C5-glutamine.
Cells, ex vivo tissues, animal models and patients can all be incubated or infused with the labelled nutrient before samples are extracted for analysis of 13C incorporation into intracellular and extracellular metabolites . Using these methods, we can directly assess activity of enzymes within diverse metabolic pathways, whose activity can change significantly without alterations in the transcriptome or proteome (e.g. Hollinshead et al. Cell Reports 2018; Lussey et al. Nat Commun 2016).
Our analyses are further enhanced by the combined use of mass spectrometry and NMR spectroscopy methods, which together provide both positional information on the incorporation of 13C atoms into metabolites as well as high sensitivity, which can be critical for unpicking similar metabolic pathways. Our investigations of cell metabolism are complemented by use of two Oroboros Oxograph-2k instruments for high resolution respirometry measurements, through which we can examine mitochondrial respiratory activity and how it alters in different conditions.
Overall, our expertise and instrumentation permits highly detailed analyses of central carbon metabolism at multiple levels – in cells, tissues and whole animals.