Scalable Phosphotyrosine Enrichment with SH2 Superbinder Enables Deep Profiling of EGF Responses

ABSTRACT: Phosphotyrosine signaling plays a critical role in many biological processes, from cell proliferation to immune response. Despite its importance, proteomic studies of tyrosine phosphorylation have been limited in scale and throughput by costly reagents and labor-intensive protocols. To address this, we developed R2HaPpY, a phosphotyrosine enrichment method combining simplified superbinder reagent preparation and automated high-throughput enrichment. Our new reagent binds phosphotyrosine peptides at higher efficiency than existing enrichment reagents and reduces both cost and preparation time by 20-fold. We generalized R2HaPpY to samples of low and high phosphotyrosine levels, and benchmarked it on EGF signaling dynamics in HeLa cells. Using only ~1 mg of input peptides, we quantify 1,651 u

SUBMITTER: Alexis Chang 

PROVIDER: S-SCDT-10_1038-S44318-026-00843-8 | biostudies-other |

REPOSITORIES: biostudies-other