Dynamic Phosphoproteomics of BRS3 Activation Reveals the Hippo Signaling Pathway for Cell Migration

• DOI: 10.1021/acs.jproteome.3c00116
• Publication Date: 2023-06-27T17:47:31Z
• Authors: Miao Guo; Yan Zhang; Lehao Wu; Ye Sheng; Jiaqi Zhao; Zeyuan Wang; Huiyu Wang; Lu Zhang; Hua Xiao
• Keywords: Phosphoproteomics; Hippo signaling pathway; Phosphopeptide; Cell Signaling

Bombesin receptor subtype-3 (BRS3) is an orphan G-protein coupled (GPCR) that involved in a variety of pathological and physiological processes, while its biological functions underlying regulatory mechanisms remain largely unknown. In this study, quantitative phosphoproteomics approach was employed to comprehensively decipher the signal transductions occurred upon intracellular BRS3 activation. The lung cancer cell line H1299-BRS3 treated with MK-5046, agonist BRS3, for different durations. Harvested cellular proteins were digested phosphopeptides enriched by immobilized titanium (IV) ion affinity chromatography (Ti4+-IMAC) label-free quantification (LFQ) analysis. A total 11,938 identified, corresponding 3,430 phosphoproteins 10,820 phosphosites. Data analysis revealed 27 six Hippo signaling pathway, which significantly regulated Verification experiments demonstrated downregulation pathway caused activation could induce dephosphorylation nucleus localization Yes-associated protein (YAP), association migration further confirmed kinase inhibition. Our data collectively demonstrate contributes through pathway.