Nat Methods. 2011 Jul 10;8(8):655-8. doi: 10.1038/nmeth.1647.

Large-scale phosphosite quantification in tissues by a spike-in SILAC method.

Monetti M, Nagaraj N, Sharma K, Mann M.

Max Planck Institute for Biochemistry, Department of Proteomics and Signal Trasduction, Am Klopferspitz 18, Martinsried D 82152, Germany.

 

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.

 

http://www.ncbi.nlm.nih.gov/pubmed/21743459

 

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© 2012, Max Planck Institute of Biochemistry, Martinsried