Novel Protein Kinase A-mediated Endothelial Cell Myosin Light Chain Kinase Phosphorylation Sites Using Data Dependent Nano-LC/MS/MS Mass Spectrometry Method

摘要

Protein phosphorylation plays a key role in vascular signaling and barrier regulation. We previously demonstrated that the non muscle myosin light chain kinase (nmMLCK) isoform present in endothelial cells, is a multi-functional enzyme which drives the participation of the endothelial actin cytoskeleton in vascular barrier regulation and trafficking of inflammatory cells into the lung. Our previous studies confirmed that cAMP -dependent protein kinase A (PKA) mediates nmMLCK activity via phosphorylation(1). Here we use optimized biochemical protocols and mass spectrometry methods to further map novel additional phosphorylation sites of PKA-mediated nmMLCK.