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Ubiquitin specific protease 7 (USP7) belongs to the family of deubiquitinat-ing enzymes. Among other functions, USP7 is involved in the regulation of stress response pathways, epigenetic silencing and the progress of infections by DNA viruses. | ễFEBS Journal Biochemical characterization of USP7 reveals post-translational modification sites and structural requirements for substrate processing and subcellular localization Amaury Fernandez-Montalvan1 Tewis Bouwmeester2 Gerard Joberty2 Robert Mader3 Marion Mahnke4 Benoit Pierrat1 Jean-Marc Schlaeppi4 Susanne Worpenberg1 and Bernd Gerhartz1 1 Expertise Platform Proteases Novartis Institutes for BiomedicalResearch Basel Switzerland 2 Cellzome AG Heidelberg Germany 3 MusculoskeletalDisease Area Novartis Institutes for BiomedicalResearch Basel Switzerland 4 Biologics Centre Novartis Institutes for BiomedicalResearch Basel Switzerland Keywords biochemical characterization cysteine protease deubiquitinating enzyme ubiquitin pathway USP7 HAUSP Correspondence A. Fernández-Montalván Molecular Screening and Cellular Pharmacology Merck Serono S.A. 9 Chemin des Mines Case postale 54 CH-1211 Geneva 20 Switzerland Fax 41 22 4149558 Tel 41 22 4144977 E-mail amaury.fernandez@merckserono.net B. Gerhartz Expertise Platform Proteases Novartis Institutes for Biomedical Research CH-4002 Basel Switzerland Fax 41 61696 8132 Tel 41 61696 1204 E-mail bernd.gerhartz@novartis.com Received 20 April2007 revised 14 June 2007 accepted 25 June 2007 doi 10.1111 j.1742-4658.2007.05952.x Ubiquitin specific protease 7 USP7 belongs to the family of deubiquitinat-ing enzymes. Among other functions USP7 is involved in the regulation of stress response pathways epigenetic silencing and the progress of infections by DNA viruses. USP7 is a 130-kDa protein with a cysteine peptidase core N- and C-terminal domains required for protein-protein interactions. In the present study recombinant USP7 full length along with several variants corresponding to domain deletions were expressed in different hosts in order to analyze post-translational modifications oligomerization state enzymatic properties and subcellular localization patterns of the enzyme. USP7 is phosphorylated at S18 and S963 and ubiquitinated .