title: Modelling and molecular dynamics of the interaction between the E3 ubiquitin ligase Itch and the E2 UbcH7.
abstract: Itch, a member of the E6AP carboxy terminus (HECT) domain-containing family of ubiquitin E3 ligases, acts in concert with the ubiquitin activating enzyme (E1) and the ubiquitin conjugating enzyme (E2) to catalyze ubiquitylation of protein targets. This sub-family of E3s shares a 350 residue C-terminal HECT domain having a strictly conserved catalytic Cys, and recruiting its cognate ubiquitin-loaded E2. HECT domains possess intrinsic enzymatic activity, by accepting ubiquitin from an E2, forming a ubiquitin thiolester intermediate, and directly catalyzing ubiquitylation of the target protein. Several hypotheses have been proposed for the biochemical mechanism underlying the structural relationship of the HECT-E2 association and subsequent ubiquitin transfer. Nonetheless, a detailed characterization of the process is still missing. In this work, we have used molecular dynamic simulations, free energy calculations, protein modelling techniques and normal modes analysis to get a deeper characterization of the static and dynamical properties of this interaction mechanism. We hypothesize a correlated slow-frequency motion that involves two different hinge regions of the HECT domain. The identification of the amino acid residues responsible for the HECT-E2 interaction, and for the dynamical properties of the ubiquitin transfer process, may be of relevant interest for pharmacological and therapeutical purposes.
authors.affiliation: Department of Biochemical Sciences "Rossi-Fanelli", University of Rome "La Sapienza", P.le Aldo Moro 5, Rome, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Raimondo
authors.forename: Alejandro
authors.initials: A
authors.lastname: Giorgetti
authors.forename: Francesca
authors.initials: F
authors.lastname: Bernassola
authors.forename: Gerry
authors.initials: G
authors.lastname: Melino
authors.forename: Anna
authors.initials: A
authors.lastname: Tramontano
mesh_terms: D006801:Humans; D008958:Models, Molecular; D011485:Protein Binding; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D055672:Static Electricity; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D012097:Repressor Proteins; D044763:Ubiquitin-Conjugating Enzymes; C432966:ITCH protein, human; D044767:Ubiquitin-Protein Ligases