title: Comprehensive identification of substrates for F-box proteins by differential proteomics analysis.
abstract: Although elucidation of enzyme-substrate relations is fundamental to the advancement of biology, universal approaches to the identification of substrates for a given enzyme have not been established. It is especially difficult to identify substrates for ubiquitin ligases, given that most such substrates are immediately ubiquitylated and degraded as a result of their association with the enzyme. We here describe the development of a new approach, DiPIUS (differential proteomics-based identification of ubiquitylation substrates), to the discovery of substrates for ubiquitin ligases. We applied DiPIUS to Fbxw7α, Skp2, and Fbxl5, three of the most well-characterized F-box proteins, and identified candidate substrates including previously known targets. DiPIUS is thus a powerful tool for unbiased and comprehensive screening for substrates of ubiquitin ligases.
authors.affiliation: Department of Molecular and Cellular Biology, Medical Institute of Bioregulation, Kyushu University , 3-1-1 Maidashi, Higashi-ku, Fukuoka, Fukuoka 812-8582, Japan.
authors.forename: Kanae
authors.initials: K
authors.lastname: Yumimoto
authors.forename: Masaki
authors.initials: M
authors.lastname: Matsumoto
authors.forename: Koji
authors.initials: K
authors.lastname: Oyamada
authors.forename: Toshiro
authors.initials: T
authors.lastname: Moroishi
authors.forename: Keiichi I
authors.initials: KI
authors.lastname: Nakayama
mesh_terms: D000818:Animals; D044783:F-Box Proteins / Q000302:isolation & purification / Q000378:metabolism*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D051379:Mice; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D059748:Proteolysis; D040901:Proteomics; D044843:SKP Cullin F-Box Protein Ligases / Q000378:metabolism*; D057149:Ubiquitinated Proteins / Q000302:isolation & purification / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D044783:F-Box Proteins; D057149:Ubiquitinated Proteins; D044843:SKP Cullin F-Box Protein Ligases