Structure solution of full-length NEDD4 E3 ligase

E3 enzymes (Ubiquitin ligases) are the final actors of the ubiquitination cascade and the critical determinants of substrate and chain specificity. As such, they are the perfect target for inhibitors designed to block specific cellular pathways. While functional and structural data are accumulating on the catalytic domains of E3 ligases, molecular details and regulatory mechanisms in the context of the full-length enzyme remain largely unknown. NEDD4 is the founding member of a class of HECT E3 ligases. We and others have detailed the molecular mechanism of ubiquitin transfer by NEDD4 using the isolated HECT. Little is known on the structural and functional properties of the holoenzyme. By applying to the CM01 CryoEM SOS pipeline our aim is to determine the high-resolution structure of the NEDD4 full-length wild-type enzyme to uncover the full structural determinants of its regulation and formally prove our biochemical results.