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  • Title: Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) Technology in Fission Yeast.
    Author: Maček B, Carpy A, Koch A, Bicho CC, Borek WE, Hauf S, Sawin KE.
    Journal: Cold Spring Harb Protoc; 2017 Jun 01; 2017(6):pdb.top079814. PubMed ID: 28572211.
    Abstract:
    Shotgun proteomics combined with stable isotope labeling by amino acids in cell culture (SILAC) is a powerful approach to quantify proteins and posttranslational modifications across the entire proteome. SILAC technology in Schizosaccharomyces pombe must cope with the "arginine conversion problem," in which isotope-labeled arginine is converted to other amino acids. This can be circumvented by either using stable isotope-marked lysine only (as opposed to the more standard lysine/arginine double labeling) or using yeast genetics to create strains that only very inefficiently convert arginine. Both strategies have been used successfully in large-scale (phospho)proteomics projects in S. pombe Here we introduce methods for performing a typical SILAC-based experiment in fission yeast, including generation of SILAC-compatible strains, sample preparation, and measurement by mass spectrometry.
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