Characterization of V2R Pharmacoperones

SR121463B is a V2R antagonist and known pharmacoperone that was used in the current study, after being generously provided by Dr. Claudine Serradeil at Sanofi-Aventis and used as received. Other pharmacoperones were identified by us by high throughput screening of a large chemical library [22 (link), 23 (link)]. Several reagents were used as obtained from indicated vendors: 3-Isobutyl-1-methylxanthine (IBMX, Sigma Aldrich, St. Louis, MO), vasopressin (Tocris Biosciences, Bristol, England UK), fetal calf serum (FCS, Hyclone, Logan, UT), Dulbecco’s MEM (DMEM), PBS (GIBCO, Invitrogen). pTRE2-Hygromycin vector (Invitrogen, San Diego, CA), human arginine-vasopressin 2 receptor (V2R; Gene Bank Accession Number: AY242131), Gq plasmid (Gene Bank Accession Number: U43083) [24 (link)]; both plasmids from cDNA Resource Center; www.cdna.org;), myo-[2-³H(N)]-inositol (NET-114A; PerkinElmer, Waltham, MA), vasopressin (8-L-arginine), [phenylalaninyl-3,4,5-³H(N)- (NET800, specific activity = 66.3 Ci/mmol; PerkinElmer, Waltham, MA), and unbound 125-Iodine (016303710; MP Biomedicals, Santa Ana, CA).