title: Structural basis of ubiquitin recognition by the deubiquitinating protease USP2.
abstract: Deubiquitinating proteases reverse protein ubiquitination and rescue their target proteins from destruction by the proteasome. USP2, a cysteine protease and a member of the ubiquitin specific protease family, is overexpressed in prostate cancer and stabilizes fatty acid synthase, which has been associated with the malignancy of some aggressive prostate cancers. Here, we report the structure of the human USP2 catalytic domain in complex with ubiquitin. Ubiquitin uses two major sites for the interaction with the protease. Both sites are required simultaneously, as shown by USP2 inhibition assays with peptides and ubiquitin mutants. In addition, a layer of ordered water molecules mediates key interactions between ubiquitin and USP2. As several of those molecules are found at identical positions in the previously solved USP7/ubiquitin-aldehyde complex structure, we suggest a general mechanism of water-mediated ubiquitin recognition by USPs.
authors.affiliation: Protease Platform, Novartis Institutes for BioMedical Research, 4002 Basel, Switzerland. martin.renatus@novartis.com
authors.forename: Martin
authors.initials: M
authors.lastname: Renatus
authors.forename: Shirley Gil
authors.initials: SG
authors.lastname: Parrado
authors.forename: Allan
authors.initials: A
authors.lastname: D'Arcy
authors.forename: Ulf
authors.initials: U
authors.lastname: Eidhoff
authors.forename: Bernd
authors.initials: B
authors.lastname: Gerhartz
authors.forename: Ulrich
authors.initials: U
authors.lastname: Hassiepen
authors.forename: Benoit
authors.initials: B
authors.lastname: Pierrat
authors.forename: Ralph
authors.initials: R
authors.lastname: Riedl
authors.forename: Daniela
authors.initials: D
authors.lastname: Vinzenz
authors.forename: Susanne
authors.initials: S
authors.lastname: Worpenberg
authors.forename: Markus
authors.initials: M
authors.lastname: Kroemer
mesh_terms: D000595:Amino Acid Sequence; D003460:Crystallization; D017931:DNA Primers; D010450:Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008670:Metals / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism*; D043222:Ubiquitin Thiolesterase; D014867:Water / Q000378:metabolism
chemical_list: D017931:DNA Primers; D008670:Metals; D025801:Ubiquitin; D014867:Water; D010450:Endopeptidases; C108665:USP2 protein, human; D043222:Ubiquitin Thiolesterase