Label-Free Quantitative Phosphoproteomics in the Fission Yeast Schizosaccharomyces pombe

Methods Mol Biol. 2025:2862:297-307. doi: 10.1007/978-1-0716-4168-2_21.

Abstract

Protein phosphorylation is a dynamic, reversible posttranslational modification that plays an important role in the regulation of cell signaling. Recently, label-free quantitative (LFQ) phosphoproteomics has become a powerful tool to analyze the phosphorylation of proteins within complex samples. In this chapter, we describe how to apply LFQ phosphoproteomics that is based on Fe-IMAC phosphopeptide enrichment followed by strong anion exchange (SAX) and porous graphitic carbon (PGC) fractionation strategies for identification and quantification of changes in the phosphoproteome in the fission yeast Schizosaccharomyces pombe.

Keywords: LFQ phosphoproteomics; Mass spectrometry; PGC fractionation; Phosphopeptide enrichment; SAX fractionation; Schizosaccharomyces pombe.

MeSH terms

  • Phosphopeptides / analysis
  • Phosphopeptides / metabolism
  • Phosphoproteins* / analysis
  • Phosphoproteins* / metabolism
  • Phosphorylation
  • Protein Processing, Post-Translational
  • Proteome / analysis
  • Proteome / metabolism
  • Proteomics* / methods
  • Schizosaccharomyces pombe Proteins* / metabolism
  • Schizosaccharomyces* / metabolism

Substances

  • Phosphoproteins
  • Schizosaccharomyces pombe Proteins
  • Phosphopeptides
  • Proteome