Ubiquitin-activating enzyme (E1) catalyzed amidation was developed to functionalize the ubiquitin C-terminus with various unique functional groups, such as thiol, azide, alkyne, and alkene, with high efficiency and high yield (>90%). These unique functional groups were successfully applied to undergo facile syntheses of fluorophore-conjugated ubiquitin and polyubiquitin mimics that serve as substrates of deubiquitinases. Furthermore, this method can be applied to synthesize various mixed linkage types(up to 49) of triubiquitin mimics, which can provide abundant diversified ubiquitin chains for the structural and functional study of the ubiquitin system.

E1-catalyzed ubiquitin C-terminal amidation for the facile syntheses of deubiquitinase substrates

247th National Meeting (2014)

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