Monitoring GPCR Induced Atypical MAPK p38 Signaling to Identify Key Regulators of Vascular Inflammation

Abstract

Activation of the mitogen activated protein kinase (MAPK) p38 is a critical regulatory step in pro‐inflammatory signaling and a key target for therapeutic intervention in vascular diseases, including chronic inflammation and lung disease. Our recent studies defined a conserved ubiquitin driven pathway that regulates atypical p38 signaling by a subset of G protein‐coupled receptors (GPCR). Atypical p38 signaling is initiated by GPCR induced c‐Src phosphorylation of an autoinhibitory peptide in the E3 ubiquitin ligase NEDD4‐2. In turn, phospho‐activation releases the autoinhibitory peptide from the catalytic site of NEDD4‐2, a step which is critical for stress induced GPCR ubiquitination and amplification of atypical p38 signaling and inflammation in the vasculature. Despite the critical role of GPCR induced p38 signaling in the onset and progression of many pathophysiological responses, atypical p38 signaling is still relatively understudied. Here we report the development of Försters Resonance Energy Transfer (FRET) biosensors to map the spatiotemporal activation of p38 signaling by multiple GPCRs in vascular endothelial cells. These FRET sensors create an ideal platform for a small‐scale screening pipeline to identify molecular modulators of the atypical p38 signaling pathway. Our studies have already identified multiple regulatory proteins that block the induction and/or magnitude of GPCR induced p38 signaling, perturbing pro‐inflammatory signaling in the vasculature.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.