title: A clathrin-coated vesicle-mediated pathway in atrial natriuretic peptide (ANP) secretion.
abstract: Atrial Natriuretic Peptide (ANP), produced by atrial cardiomyocytes, is an endogenous hypotensive agent that brings about vasodilation and diuresis. Similar to other polypeptide hormones, ANP is synthesized as a precursor, preproANP. The preprohormone is processed intracellularly and is stored in secretory granules as the prohormone. During the events of exocytosis, the prohormone is converted to its active form, ANP. In this study, a double-label immunocytochemistry experiment was performed using ANP and clathrin antibodies to determine if the transport of this hormone is mediated by clathrin-coated vesicles. Additionally, we have isolated clathrin-coated vesicles (CVs) from adult rat atria using immunoadsorption, and have characterized the fraction by using SDS PAGE, TEM, and Western blot analysis. The data demonstrate that: (1) ANP and clathrin co-localize in myocardial tissue, (2) clathrin-coated vesicles can be isolated from adult rat atria, and (3) clathrin-coated vesicles isolated from adult atrial myocardium contain predominantly proANP. The presence of proANP in clathrin-coated vesicles suggests that this polypeptide hormone is transported intracellularly via a clathrin-mediated pathway and during transit the prohormone is not significantly converted to its active form.
authors.affiliation: Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City 66160-7400.
authors.forename: R M
authors.initials: RM
authors.lastname: Klein
authors.forename: K B
authors.initials: KB
authors.lastname: Kelley
authors.forename: E M
authors.initials: EM
authors.lastname: Merisko-Liversidge
mesh_terms: D000818:Animals; D009320:Atrial Natriuretic Factor / Q000378:metabolism*; D001692:Biological Transport; D002458:Cell Fractionation; D002966:Clathrin / Q000032:analysis*; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000378:metabolism; D005089:Exocytosis; D006325:Heart Atria / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D002966:Clathrin; D011498:Protein Precursors; D009320:Atrial Natriuretic Factor