Subcellular trafficking of guanylyl cyclase/natriuretic peptide receptor-A with concurrent generation of intracellular cGMP

摘要: Atrial natriuretic peptide (ANP) activates guanylyl cyclase/natriuretic receptor-A (GC-A/NPRA), which lowers blood pressure and volume. The objective of the present study was to visualize internalization trafficking enhanced GFP (eGFP)-tagged NPRA (eGFP–NPRA) in human embryonic kidney-293 (HEK-293) cells, using immunofluorescence (IF) co-immunoprecipitation (co-IP) eGFP–NPRA. Treatment cells with ANP initiated rapid co-localization receptor early endosome antigen-1 (EEA-1), highest at 5 min gradually decreased within 30 min. Similarly, observed lysosome-associated membrane protein-1 (LAMP-1); however, after treatment lysosomotropic agents, intracellular accumulation increased Co-IP assays confirmed that localization internalized receptors occurred subcellular organelles during endocytosis NPRA. Rab 11, used as a recycling (Re) marker, indicated ∼20% recycled back plasma membrane. ANP-treated showed marked increase IF cGMP, whereas still into compartments. Thus, ligand binding, is rapidly trafficked from cell surface endosomes, Res lysosomes, concurrent generation cGMP.