Detection of metabolic activity enables
us to reveal
the inherent
metabolic state of cells and elucidate mechanisms underlying cellular
homeostasis and growth. However, a fluorescence approach for the study
of metabolic pathways is still largely unexplored. Herein, we have
developed a new chemical probe for the fluorescence-based detection
of fatty acid β-oxidation (FAO), a key process in lipid catabolism,
in cells and tissues. This probe serves as a substrate of FAO and
forms a reactive quinone methide (QM) as a result of metabolic reactions.
The liberated QM is covalently captured by intracellular proteins,
and subsequent bio-orthogonal ligation with a fluorophore enables
fluorescence analysis. This reaction-based sensing allowed us to detect
FAO activity in cells at a desired emission wavelength using diverse
analytical techniques including fluorescence imaging, in-gel fluorescence
activity-based protein profiling (ABPP), and fluorescence-activated
cell sorting (FACS). The probe was able to detect changes in FAO activity
induced by chemical modulators in cultured cells. The probe was further
employed for fluorescence imaging of FAO in mouse liver tissues and
revealed the metabolic heterogeneity of FAO activity in hepatocytes
by the combination of FACS and gene expression analysis, highlighting
the utility of our probe as a chemical tool for fatty acid metabolism
research.
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