title: Heterologous downregulation of vasopressin type 2 receptor is induced by transferrin.
abstract: Vasopressin (VP) binds to the vasopressin type 2 receptor (V2R) to trigger physiological effects including body fluid homeostasis and blood pressure regulation. Signaling is terminated by receptor downregulation involving clathrin-mediated endocytosis and V2R degradation. We report here that both native and epitope-tagged V2R are internalized from the plasma membrane of LLC-PK1 kidney epithelial cells in the presence of another ligand, transferrin (Tf). The presence of iron-saturated Tf (holo-Tf; 4 h) reduced V2R binding sites at the cell surface by up to 33% while iron-free (apo-Tf) had no effect. However, no change in green fluorescent protein-tagged V2R distribution was observed in the presence of bovine serum albumin, atrial natriuretic peptide, or ANG II. Conversely, holo-Tf did not induce the internalization of another G protein-coupled receptor, the parathyroid hormone receptor. In contrast to the effect of VP, Tf did not increase intracellular cAMP or modify aquaporin-2 distribution in these cells, although addition of VP and Tf together augmented VP-induced V2R internalization. Tf receptor coimmunoprecipitated with V2R, suggesting that they interact closely, which may explain the additive effect of VP and Tf on V2R endocytosis. Furthermore, Tf-induced V2R internalization was abolished in cells expressing a dominant negative dynamin (K44A) mutant, indicating the involvement of clathrin-coated pits. We conclude that Tf can induce heterologous downregulation of the V2R and this might desensitize VP target cells without activating downstream V2R signaling events. It also provides new insights into urine-concentrating defects observed in rat models of hemochromatosis.
authors.affiliation: 1Nephrology Division, MGH Center for Systems Biology, Program in Membrane Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114, USA. bouley.richard@mgh.harvard.edu
authors.forename: Richard
authors.initials: R
authors.lastname: Bouley
authors.forename: Paula
authors.initials: P
authors.lastname: Nunes
authors.forename: Billy
authors.initials: B
authors.lastname: Andriopoulos
authors.forename: Margaret
authors.initials: M
authors.lastname: McLaughlin
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Webber
authors.forename: Herbert Y
authors.initials: HY
authors.lastname: Lin
authors.forename: Jodie L
authors.initials: JL
authors.lastname: Babitt
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Gardella
authors.forename: Dennis A
authors.initials: DA
authors.lastname: Ausiello
authors.forename: Dennis
authors.initials: D
authors.lastname: Brown
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects*; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D017483:Receptors, Vasopressin / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013552:Swine; D014168:Transferrin / Q000494:pharmacology*; D014667:Vasopressins / Q000494:pharmacology*
chemical_list: D017483:Receptors, Vasopressin; D014168:Transferrin; D014667:Vasopressins