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


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.


Projects: A1.3: Identification of crucial metabolic processes during hepatocyte pr..., C5: Structural changes and functional consequences of the liver lobular/..., HepatoSys

Nat. Methods
Nat. Methods 8(8): 655-8
10th Jul 2011

Mara Monetti, Nagarjuna Nagaraj, Kirti Sharma, Matthias Mann

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[Mara Monetti] [Kirti Sharma] [Matthias Mann]

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  • Created: 30th Jul 2012 at 13:35
  • Last updated: 24th Oct 2013 at 16:18

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