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Keywords:

  • deubiquitinases;
  • diubiquitin;
  • protein engineering;
  • synthetic biology;
  • ubiquitin functionalization
Thumbnail image of graphical abstract

Will Ub my partner? The ubiquitin (Ub)-activating enzyme (E1) was used to catalyze an amidation reaction to functionalize the C terminus of Ub with unique functional groups, such as thiol, azide, alkyne, and alkene groups, with high efficiency and yield (>90 %). These groups were then applied for the facile synthesis of fluorophore-conjugated ubiquitin and specifically conjugated diubiquitin substrates for deubiquitinases.