A Fluorogenic Phosphine-Ester Conjugate for Monitoring Protein S-nitrosylation in Endothelial Cells
【摘要】：As a post-translational modification, protein S-nitrosylation has been increasingly recognized as an important signaling process in vascular physiology and pathology, and its in situ detection would provide valuable information to interrogate the molecular mechanisms underpinning related-diseases. Motivated by the dearth of chemical tools capable of directly imaging protein S-nitrosylation in biological contexts, we have developed a fluorogenic probe based on the phosphine-RSNO chemistry. The probe is weakly emissive alone and becomes highly fluorescent only after undergoing reaction with S-nitrosothiols in live cellular environments. It has been successfully applied to image the dynamic changes of protein S-nitrosylation in live endothelial cells. Its specificity in complex biological systems has been further verified by imaging a known target of S-nitrosylation, glyceraldehyde-3-phosphate dehydrogenase(GAPDH), in live cells.