title: Binding, internalization, and degradation of atrial natriuretic peptide in cultured vascular smooth muscle cells of rat.
abstract: Binding, internalization, and degradation of 125I-labeled-rat atrial natriuretic peptide (rANP) were studied in cultured rat aortic vascular smooth muscle cells (VSMC). At 37 degrees C, 125I-labeled-rANP rapidly bound to VSMCs, but the cell-bound radioactivity rapidly decreased upon subsequent incubation, while the binding was slow at 4 degrees C, reaching to an apparent equilibrium after 6 hrs. The cell-bound 125I-labeled-rANP at 37 degrees C is rapidly dissociated from VSMC (t 1/2: approximately 40 min) with the appearance of degradaded product(s) of radioligand in the medium, whereas the degradation was minimal at 4 degrees C. This degradative process was blocked by inhibitors of metabolic energy production (azide, dinitrophenol), inhibitors of lysosomal cathepsins (leupeptin, pepstatin), and lysosomotropic agents (NH4Cl, chloroquine, lidocaine, methylamine, dansylcadaverine), but not by inhibitors of serine or thiol proteases. 125I-labeled-rANP initially bound to the cell-surface was rapidly internalized, and delivered to lysosomal structures, which was confirmed by autoradiographic studies. These data indicate that rANP, after binding to the cell-surface receptors, is rapidly internalized into the cells through receptor-mediated endocytosis, and subsequently degradaded by lysosomal hydrolases.
authors.forename: Y
authors.initials: Y
authors.lastname: Hirata
authors.forename: S
authors.initials: S
authors.lastname: Takata
authors.forename: M
authors.initials: M
authors.lastname: Tomita
authors.forename: S
authors.initials: S
authors.lastname: Takaichi
mesh_terms: D000818:Animals; D009320:Atrial Natriuretic Factor / Q000378:metabolism*; D001345:Autoradiography; D002478:Cells, Cultured; D004705:Endocytosis; D007457:Iodine Radioisotopes; D008247:Lysosomes / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D051381:Rats; D013696:Temperature; D011503:Transglutaminases / Q000502:physiology
chemical_list: D007457:Iodine Radioisotopes; D009320:Atrial Natriuretic Factor; D011503:Transglutaminases