title: High-throughput screen for pharmacoperones of the vasopressin type 2 receptor.
abstract: Pharmacoperone drugs correct the folding of misfolded protein mutants and restore function (i.e., "rescue") by correcting the routing of (otherwise) misrouted mutants. Assays for pharmacoperones have not been applied to screen large libraries previously. Currently, most pharmacoperones possess intrinsic agonist or antagonist activities since these were identified using high-throughput screens aimed at discovering direct agonists or antagonists. Here we describe an ultra-high-throughput compatible no-wash assay system designed to specifically identify pharmacoperones of the vasopressin type 2 receptor (V2R). Development of such assays is important and novel since useful chemical structures with the ability to control cellular trafficking but lacking intrinsic agonist or antagonist properties have not likely been identified using existing screens. In the described assay, the level of functional human V2R (hV2R) (mutant) present in each test well is quantitated by stimulation with saturating levels of agonist followed by use of a luminescent-based cyclic adenosine monophosphate assay. This allows the assay to identify compounds that increase the trafficking of mutant hV2R[L(83)Q] in our model system.
authors: Divisions of Reproductive Sciences and Neuroscience, Oregon National Primate Research Center, Beaverton, OR 97006, USA. connm@ohsu.edu
authors: P Michael
authors: PM
authors: Conn
authors: Emery
authors: E
authors: Smith
authors: Peter
authors: P
authors: Hodder
authors: Jo Ann
authors: JA
authors: Janovick
authors: David C
authors: DC
authors: Smithson
mesh_terms: D055808:Drug Discovery; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D017510:Protein Folding / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects*; D057165:Proteostasis Deficiencies / Q000188:drug therapy*; D017483:Receptors, Vasopressin / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism
chemical_list: C585768:AVPR2 protein, human; D017483:Receptors, Vasopressin
keywords: GPCRs; chemical library; pharmacoperone; protein; vasopressin type 2 receptor (V2R)
references: Pharmacol Rev. 2007 Sep;59(3):225-50
references: 17878512
references: J Pharmacol Exp Ther. 2003 May;305(2):608-14
references: 12606630
references: Endocr Rev. 2005 Jun;26(4):479-503
references: 15536207
references: J Clin Endocrinol Metab. 2002 Jul;87(7):3255-62
references: 12107234
references: J Clin Invest. 2000 Apr;105(7):887-95
references: 10749568
references: Trends Pharmacol Sci. 2009 May;30(5):228-33
references: 19307028
references: J Am Soc Nephrol. 2006 Jan;17(1):232-43
references: 16319185
references: J Am Soc Nephrol. 1996 Mar;7(3):410-4
references: 8704106
references: Mol Cell Endocrinol. 2007 Jun 30;272(1-2):77-85
references: 17555869
references: Methods Enzymol. 2013;521:3-16
references: 23351731
references: Cardiovasc Drug Rev. 2001 Fall;19(3):201-14
references: 11607038
references: Trends Endocrinol Metab. 2010 Mar;21(3):190-7
references: 20005736
references: Anal Biochem. 2003 Feb 15;313(2):246-54
references: 12605861
references: Assay Drug Dev Technol. 2007 Apr;5(2):237-45
references: 17477832
references: PLoS One. 2011;6(7):e22784
references: 21818389