title: Importance of the mercury-sensitive cysteine on function and routing of AQP1 and AQP2 in oocytes.
abstract: To discriminate between water transport of of aquaporin-2 (AQP2) mutants in nephrogenic diabetes insipidus and that of an AQP2 molecule used to drag them to the oolemma, we investigated the mercury sensitivity of wild-type and AQP2 C181S proteins in oocytes. Incubation with HgCl2 inhibited the osmotic water permeability (Pf) of human (h) AQP2 by 40%, whereas inhibition of hAQP1 was 75%. Oocytes expressing hAQP1 C189S revealed a Pf comparable to wild-type hAQP1, but mercury sensitivity was lost. In contrast, no increase in Pf was obtained when hAQP2 C181S was expressed. Also, expression of rat AQP2 C181A and C181S mutants did not increase the Pf, which contrasts with published observations. Immunocytochemistry and immunoblotting revealed that only AQP1, AQP1 C189S, and AQP2 were targeted to the plasma membrane and that AQP2 mutant proteins are retarded in the endoplasmic reticulum. In conclusion, water transport through AQP2 is less sensitive to mercury inhibition than through AQP1. Furthermore, substitution of the mercury-sensitive cysteine for a serine results in an impaired routing of human and rat AQP2. Similar mutations have no effect on AQP1 function, which is indicative of structural differences between AQP1 and AQP2.
authors.affiliation: Department of Cell Physiology, University of Nijmegen, The Netherlands.
authors.forename: S M
authors.initials: SM
authors.lastname: Mulders
authors.forename: J P
authors.initials: JP
authors.lastname: Rijss
authors.forename: A
authors.initials: A
authors.lastname: Hartog
authors.forename: R J
authors.initials: RJ
authors.lastname: Bindels
authors.forename: C H
authors.initials: CH
authors.lastname: van Os
authors.forename: P M
authors.initials: PM
authors.lastname: Deen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D051398:Aquaporin 1; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins; D001789:Blood Group Antigens; D002463:Cell Membrane Permeability; D017124:Conserved Sequence; D003545:Cysteine; D003919:Diabetes Insipidus / Q000235:genetics / Q000503:physiopathology; D005260:Female; D006801:Humans; D007473:Ion Channels / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D008627:Mercuric Chloride / Q000494:pharmacology*; D008961:Models, Structural; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D017354:Point Mutation; D017433:Protein Structure, Secondary; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000187:drug effects; D014158:Transcription, Genetic; D014982:Xenopus laevis
chemical_list: C496020:AQP1 protein, human; C496023:AQP2 protein, human; C496022:Aqp1 protein, rat; C496025:Aqp2 protein, rat; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins; D001789:Blood Group Antigens; D007473:Ion Channels; D011994:Recombinant Proteins; D051398:Aquaporin 1; D008627:Mercuric Chloride; D003545:Cysteine