title: Identification and application of NEDD8 E1 inhibitors.
abstract: The NEDD8 conjugation pathway is initiated by the NEDD8 E1, also known as NEDD8 activating enzyme (NAE) or APPBP1/UBA3 (Gong, Yeh. J Biol Chem 274:12063-12042, 1999). The best described biological role for NEDD8 conjugation is to regulate the activity of the cullin RING ligase (CRL) family of ubiquitin E3 ligases (Gong, Yeh. J Biol Chem 274:12063-12042, 1999). In this way, the NEDD8 pathway regulates the turnover of a subset of ubiquitin proteasome system (UPS) substrates that are essential for cancer cell growth and survival (Soucy, Smith, Milhollen. Nature 458:732-737, 2009). We recently initiated clinical trials with a first-in-class small molecule inhibitor of NAE for the treatment of cancer (Soucy, Smith, Milhollen. Nature 458:732-737, 2009). Here we describe a biochemical and cell-based assay used to identify NAE inhibitors and monitor inhibition of the NEDD8 conjugation pathway.
authors.affiliation: Discovery, Millennium Pharmaceuticals Inc., Cambridge, MA, USA.
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Bruzzese
authors.forename: Michael A
authors.initials: MA
authors.lastname: Milhollen
authors.forename: James M
authors.initials: JM
authors.lastname: Gavin
authors.forename: Helen R
authors.initials: HR
authors.lastname: Josephine
authors.forename: James E
authors.initials: JE
authors.lastname: Brownell
mesh_terms: D045744:Cell Line, Tumor; D044842:Cullin Proteins / Q000378:metabolism; D004789:Enzyme Activation; D006801:Humans; D000075687:NEDD8 Protein; D044764:Ubiquitin-Activating Enzymes / Q000037:antagonists & inhibitors*; D014452:Ubiquitins / Q000032:analysis / Q000378:metabolism*
chemical_list: D044842:Cullin Proteins; D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; D014452:Ubiquitins; D044764:Ubiquitin-Activating Enzymes