Bombesin-like receptor 3 (BRS3), an orphan G protein–coupled receptor (GPCR), plays important roles in our biological system while the exact mechanisms behind are less known. To get insights of biological effects on BRS3 activation, we utilized quantitative proteomics approach to explore the dynamic protein profiling during its stimulus. At different time points after stimulation, the protein profiling in HEK 293 cells with overexpressed BRS3 (HEK 293-BRS3) were analyzed by nano-LC−MS/MS.