title: Large-scale phosphosite quantification in tissues by a spike-in SILAC method.
abstract: Despite progress in mass spectrometry (MS)-based phosphoproteomics, large-scale in vivo analyses remain challenging. Here we report a 'spike-in' stable-isotope labeling with amino acids in cell culture (SILAC) methodology using standards derived from labeled mouse liver cell lines, using which we analyzed insulin signaling. With this approach we identified 15,000 phosphosites and quantitatively compared 10,000 sites in response to insulin treatment, creating a very large, accurately quantified in vivo phosphoproteome dataset.
authors: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors: Mara
authors: M
authors: Monetti
authors: Nagarjuna
authors: N
authors: Nagaraj
authors: Kirti
authors: K
authors: Sharma
authors: Matthias
authors: M
authors: Mann
mesh_terms: D002460:Cell Line; D002478:Cells, Cultured; D007328:Insulin / Q000494:pharmacology*; D007553:Isotope Labeling / Q000379:methods*; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D017905:Phosphites / Q000378:metabolism*
chemical_list: D007328:Insulin; D017905:Phosphites