Multiplexed Quantitative Cell Surface Proteomics by Combination of TMT Labeling and Biotinylated Peptide Enrichment

We examined changes in surface proteome of HeLa cells stimulated with epidermal growth factor (EGF), which is well known to induce the internalization and degradation of EGF receptor (EGFR). Surface biotinylation using Sulfo-NHS-biotin was performed at four different times after EGF stimulation in multiple biological replicates, followed by digestion of cellular proteins with trypsin. After TMT labeling and mixing of digested peptides, biotinylated peptides were enriched using Tamavidin 2-REV, followed by LC-MS/MS analysis.