BRAF is a serine/threonine protein kinase activating the MAP kinase/ERK-signaling pathway.Mutations of BRAF,over 90% V600E,can be detected in a wide range of human cancers.Better understanding of oncogenesis in BRAF mutated cells leads more efficient therapy for these cancers.Bioorthogonal cleavage reaction between TCOK and Me2Tz has been reporthed for kinase activation in situ.Herein,we combined this strategy and phosphoproteomics to investigate how BRAFV600E protein induces tumorigenesis.