کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1214719 | 966945 | 2009 | 8 صفحه PDF | دانلود رایگان |
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 14-3-3ɛ, the isoform with least characterized protein interactions in 14-3-3 family in mammalian cells. Among the 19 proteins interacting with 14-3-3ɛ identified, 6 of them including SKb1Hs, p54nrb, serine/threonine kinase 38, MEP50, 14-3-3θ and cofilin 2 were the previously unknown interacting partners with 14-3-3ɛ. 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.
Journal: Journal of Chromatography B - Volume 877, Issue 7, 1 March 2009, Pages 627–634