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Using a modified single-molecule pull-down (SiMPull) approach, the first direct detection of activation-dependent multisite phosphorylation on intact EGFR is provided. Integrating SiMPull data with rule-based modeling revealed roles for receptor dimerization dynamics and adaptor protein concentrations in directing EGFR signaling.

Multisite EGFR phosphorylation is regulated by adaptor protein abundances and dimer lifetimes