Tissue Extracts for Quantitative Mass Spectrometry of Planarian Proteins Using SILAC
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SILAC (stable isotope labeling by amino acids in cell culture) proteomics enables the relative quantification of proteins in one or more biological samples by mass spectrometry. This technology is based on the metabolic incorporation of heavy isotope-labeled essential amino acids into nascent proteins in vivo. Here, we describe the preparation of SILAC protein samples from planarians, flatworms with high regenerative potential and tissue plasticity. Applications for SILAC proteomics of planarians include the analysis of protein abundances, protein–protein interactions and turnover rates during stem cell-based regeneration and tissue homeostasis.
Key wordsPlanarians SILAC Quantitative proteomics Protein extraction Protein processing FASP
We thank S. Zanivan and M. Mann for sharing advice and SILAC mouse liver. This work was supported by the Max Planck Society and the Deutsche Forschungsgemeinschaft (SFB629, PAK479). The Bartscherer lab is part of the Cells-in-Motion Cluster of Excellence (EXC 1003-CiM) and is funded by the European Research Council (ERC-2016-StG 716894 IniReg). We thank J. Rink for helpful comments on the manuscript.
- 14.Zanivan S, Meves A, Behrendt K, Schoof EM, Neilson LJ, Cox J, Tang HR, Kalna G, van Ree JH, van Deursen JM, Trempus CS, Machesky LM, Linding R, Wickström SA, Fässler R, Mann M (2013) In vivo SILAC-based proteomics reveals phosphoproteome changes during mouse skin carcinogenesis. Cell Rep 3(2):552–566CrossRefGoogle Scholar
- 21.Wang Y, Yang F, Gritsenko MA, Wang Y, Clauss T, Liu T, Shen Y, Monroe ME, Lopez-Ferrer D, Reno T, Moore RJ, Klemke RL, Camp DG 2nd, Smith RD (2011) Reversed-phase chromatography with multiple fraction concatenation strategy for proteome profiling of human MCF10A cells. Proteomics 11(10):2019–2026CrossRefGoogle Scholar