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Internalization Of Guanylyl Cyclase/Natriuretic Peptide Receptor‐A Occurs Involving Clathrin‐Dependent Mechanisms
Author(s) -
Somanveen K,
Pandey Amitabh C,
Wu Edwin,
Arise Kiran K,
Pandey Kailash N
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.853.5
Subject(s) - internalization , clathrin , hek 293 cells , atrial natriuretic peptide , receptor , microbiology and biotechnology , transfection , endocytosis , chemistry , npr1 , natriuretic peptide , medicine , endocrinology , biology , biochemistry , heart failure , gene
Atrial and brain natriuretic peptides (ANP and BNP) bind to guanylyl‐cyclase/natriuretic peptide receptor‐A (GC‐A/NPRA) and produce second messenger cGMP. The aim of this study was to determine whether GC‐A/NPRA is internalized involving clathrin‐coated pits. Human embryonic kidney‐293 (HEK‐293) cells were stably transfected with GC‐A/NPRA cDNA and receptor internalization studies were performed utilizing 125 I‐ANP binding assay and immunoflurescence staining in NPRA‐transfected cells. Cells were treated with chlorpromazine and monodansylcadaverine, which disrupt the formation of clathrin‐coated vesicles. In the inhibitor‐treated recombinant HEK‐293 cells, 125 I‐ANP/receptor complexes were internalized only by 10–15% compared with untreated control cells. A major portion of 125 I‐ANP radioactivity was localized on the cell surface in both chloropromazine‐ and monodansylcadaverine‐treated cells, which accounted 85–90% compared with control cells. Immunoflourescence staining was utilized to confirm the endocytosis of ANP/NPRA complexes. The results demonstrated that the internalization of GC‐A/NPRA occurs involving clathrin‐mediated endocytotic mechanisms. The present findings provide important information towards the understanding the GC‐A/NPRA signaling mechanism, which is critical in the regulation of blood pressure and cardiovascular homeostasis.