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GenParts™ DNA Fragments> | … The Pup fragments ( Acel PupE/Q ΔN43 ) used for crystallization were synthesized by GenScript, while Cglu PupE or Cglu PupM was expressed and purified as described previously 9 with the addition of an anion exchange step to remove co-purified E. coli Adk. Briefly, Pup … | Get A Quote |
Pupylation is the post-translational modification of lysine side chains with prokaryotic ubiquitin-like protein (Pup) that targets proteins for proteasomal degradation in mycobacteria and other members of Actinobacteria. Pup ligase PafA and depupylase Dop are the two enzymes acting in this pathway. Although they share close structural and sequence homology indicative of a common evolutionary origin, they catalyze opposing reactions. Here, we report a series of high-resolution crystal structures of Dop in different functional states along the reaction pathway, including Pup-bound states in distinct conformations. In combination with biochemical analysis, the structures explain the role of the C-terminal residue ... More