title: Water channels encoded by mutant aquaporin-2 genes in nephrogenic diabetes insipidus are impaired in their cellular routing.
abstract: Congenital nephrogenic diabetes insipidus is a recessive hereditary disorder characterized by the inability of the kidney to concentrate urine in response to vasopressin. Recently, we reported mutations in the gene encoding the water channel of the collecting duct, aquaporin-2 (AQP-2) causing an autosomal recessive form of nephrogenic diabetes insipidus (NDI). Expression of these mutant AQP-2 proteins (Gly64Arg, Arg187Cys, Ser216Pro) in Xenopus oocytes revealed nonfunctional water channels. Here we report further studies into the inability of these missense AQP-2 proteins to facilitate water transport in Xenopus oocytes. cRNAs encoding the missense AQPs were translated with equal efficiency as cRNAs encoding wild-type AQP-2 and were equally stable. Arg187Cys AQP2 was more stable and Gly6-4Arg and Ser216Pro AQP2 were less stable when compared to wild-type AQP2 protein. On immunoblots, oocytes expressing missense AQP-2 showed, besides the wild-type 29 kDa band, an endoplasmic reticulum-retarded form of AQP-2 of approximately 32 kD. Immunoblots and immunocytochemistry demonstrated only intense labeling of the plasma membranes of oocytes expressing wild-type AQP-2. Therefore, we conclude that in Xenopus oocytes the inability of Gly64-Arg, Arg187Cys or Ser216Pro substituted AQP-2 proteins to facilitate water transport is caused by an impaired routing to the plasma membrane.
authors.affiliation: Department of Cell Physiology, University of Nijmegen, The Netherlands.
authors.forename: P M
authors.initials: PM
authors.lastname: Deen
authors.forename: H
authors.initials: H
authors.lastname: Croes
authors.forename: R A
authors.initials: RA
authors.lastname: van Aubel
authors.forename: L A
authors.initials: LA
authors.lastname: Ginsel
authors.forename: C H
authors.initials: CH
authors.lastname: van Os
mesh_terms: D000818:Animals; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins; D015152:Blotting, Northern; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002463:Cell Membrane Permeability; D018500:Diabetes Insipidus, Nephrogenic / Q000235:genetics*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005260:Female; D005808:Genes, Recessive; D006801:Humans; D007150:Immunohistochemistry; D007473:Ion Channels / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D009865:Oocytes / Q000502:physiology; D017354:Point Mutation; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D014981:Xenopus
chemical_list: C496023:AQP2 protein, human; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins; D007473:Ion Channels; D012333:RNA, Messenger