Metabolic labeling with unnatural sugar substrates is a powerful alternative for investigating glycoconjugates in living organisms. Trehalose glycolipids can be metabolically labeled with 6-azide-modified trehalose (TreAz) analogues in live mycobacteria, enabling bioorthogonal ligation with alkyne-functionalized fluorescent probes. This strategy can be used for imaging glycolipid distribution, trafficking, and dynamics as well as metabolite profiling and discovery.
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