(790.18) Crystal Structures of an E1-E2-ubiquitin Thioester Mimetic Reveal Molecular Mechanisms of Transthioesterification

Poster Board Number: A74

Shaun Olsen (University of Texas Health Science Center at San Antonio)

E1 enzymes function as gatekeepers of ubiquitin (Ub) signaling by catalyzing activation and transfer of Ub to tens of cognate E2 conjugating enzymes in a process called E1-E2 transthioesterification.  The molecular mechanisms of transthioesterification and the overall architecture of the E1-E2-Ub complex during catalysis are unknown.  Here, we determined the structure of a covalently trapped E1-E2-ubiquitin thioester mimetic.  Two distinct architectures of the complex are observed, one in which the Ub thioester (Ub(t)) contacts E1 in an open conformation and another in which Ub(t) instead contacts E2 in a dramatically different closed conformation.  Altogether our structural and biochemical data suggest that these two conformational states represent snapshots of the E1-E2-Ub complex pre- and post-thioester transfer, and are consistent with a model in which catalysis is enhanced by a Ub(t)-mediated ‘affinity switch’ that drives the reaction forward by promoting productive complex formation or substrate release depending on the conformational state.

Support or Funding Information

Research reported in this publication was supported by the NIH R01 GM115568, R01 GM128731 and CPRIT RR200030.

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