Microscale Tracing of Protein-Protein Interaction Dynamics in Cell Signaling Pathways: A Case Study on ERK1/2 Phosphorylation Mechanisms

Abstract

This study investigates the intricacies of protein-protein interactions (PPIs) within cell signaling pathways, focusing specifically on the extracellular signal-regulated kinase (ERK1/2) phosphorylation mechanisms. By employing advanced fluorescence resonance energy transfer (FRET) and mass spectrometry techniques, we quantitatively mapped the interaction network among key signaling proteins in human epithelial cells. A novel dual-color FRET analysis was utilized to reveal real-time dynamics of ERK1/2 activation. Our findings indicate a previously unidentified modulation of ERK1/2 activity by the scaffold protein KSR1, which enhances signaling efficiency by 25% and alters downstream gene expression profiles significantly. This research not only elucidates the complex interplay of cellular signaling but also highlights the potential of targeted molecular interventions in therapeutic contexts, paving the way for future studies to explore these interactions in greater detail.