Shotgun proteomics allows for the comprehensive analysis of proteins extracted from plant cells, subcellular organelles, and membranes. Previously, two-dimensional gel electrophoresis-based proteomics was used for mass spectrometric analysis of plasma membrane proteins. However, this method is not fully applicable for highly hydrophobic proteins with multiple transmembrane domains. In order to solve this problem, we here describe a shotgun proteomics method using nano-LC-MS/MS for proteins in the plasma membrane and plasma membrane microdomain fractions. The results obtained are easily applicable to label-free protein semiquantification.
Keywords: Detergent-resistant membrane; In-gel digestion; In-solution digestion; Label-free semiquantification; Microdomain; Nano-LC-MS/MS; Plasma membrane; Shotgun proteomics.