title: Analysis of the protein complex associated with 14-3-3 epsilon by a deuterated-leucine labeling quantitative proteomics strategy.
abstract: By using an unambiguous in vivo deuterated-leucine labeling quantitative proteomic approach, at close to the physiologically relevant level, we systematically profiled multiple proteins interacting with 14-3-3epsilon, the isoform with least characterized protein interactions in 14-3-3 family in mammalian cells. Among the 19 proteins interacting with 14-3-3epsilon identified, 6 of them including SKb1Hs, p54nrb, serine/threonine kinase 38, MEP50, 14-3-3theta and cofilin 2 were the previously unknown interacting partners with 14-3-3epsilon. The newly identified interactor cofilin 2 was also validated in co-transfection and co-immunoprecipitation. In contrast, with the same stringent criteria only three known partners were identified by conventional tandem affinity purification (TAP) approach. Therefore the 'in-spectra' quantitative marker of deuterated-leucine assisted to precisely identify those genuine interacting partners with minimum requirement of validation using other molecular approaches.
authors.affiliation: Department of Chemistry and Institute of Biomedical Science, Fudan University, Shanghai 200433, China; State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Shufang
authors.initials: S
authors.lastname: Liang
authors.forename: Yanbao
authors.initials: Y
authors.lastname: Yu
authors.forename: Pengyuan
authors.initials: P
authors.lastname: Yang
authors.forename: Sheng
authors.initials: S
authors.lastname: Gu
authors.forename: Yan
authors.initials: Y
authors.lastname: Xue
authors.forename: Xian
authors.initials: X
authors.lastname: Chen
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism*; D002460:Cell Line; D003903:Deuterium / Q000378:metabolism*; D006801:Humans; D007930:Leucine / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D013194:Staining and Labeling
chemical_list: D048948:14-3-3 Proteins; D003903:Deuterium; D007930:Leucine