+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

A phenotypic high throughput screening assay for the identification of pharmacoperones for the gonadotropin releasing hormone receptor



A phenotypic high throughput screening assay for the identification of pharmacoperones for the gonadotropin releasing hormone receptor



Assay and Drug Development Technologies 12(4): 238-246



We describe a phenotypic high throughput screening (HTS) calcium flux assay designed to identify pharmacoperones for the gonadotropin releasing hormone receptor (GnRHR). Pharmacoperones are target-specific, small molecules that diffuse into cells, rescue misfolded protein mutants, and restore them to function. Rescue is based on correcting the trafficking of mutants that would otherwise be retained in the endoplasmic reticulum and unable to function correctly. This approach identifies drugs with a significant degree of novelty, relying on cellular mechanisms that are not currently exploited. Development of such assays is important, since the extensive use of agonist/antagonist screens alone means that useful chemical structures may be present in existing libraries but have not been previously identified using existing methods. Our assay utilizes cell lines stably expressing a GnRHR mutant under the control of a tetracycline (OFF) transactivator. This allows us to quantitate the level of functional and properly trafficked G protein coupled receptors present in each test well. Furthermore, since we are able to turn receptor expression on and off, we can rapidly eliminate the majority of false positives from our screening results. Our data show that this approach is likely to be successful in identifying hits from large chemical libraries.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 051209240

Download citation: RISBibTeXText

PMID: 24831790

DOI: 10.1089/adt.2014.576


Related references

High-throughput screening using beta-lactamase reporter-gene technology for identification of low-molecular-weight antagonists of the human gonadotropin releasing hormone receptor. Assay and Drug Development Technologies 3(2): 143-154, 2005

Gonadotropin releasing hormone stimulation of gonadotropin release from the pituitary regulation of cell responses and gonadotropin releasing hormone receptors by gonadotropin releasing hormone receptor micro aggregation. Journal of Cellular Biochemistry Suppl. (8 Part A): 233, 1984

Identification and characterization of the gonadotropin-releasing hormone response elements in the mouse gonadotropin-releasing hormone receptor gene. Journal of Biological Chemistry 274(2): 867-880, 1999

Negative regulation of gonadotropin-releasing hormone and gonadotropin-releasing hormone receptor gene expression by a gonadotropin-releasing hormone agonist in the rat hypothalamus. Journal of Neuroendocrinology 11(3): 195-201, 1999

Identification of novel selective P2Y 6 receptor antagonists by high-throughput screening assay. Life Sciences 180: 137-142, 2017

Identification of novel human high-density lipoprotein receptor Up-regulators using a cell-based high-throughput screening assay. Journal of Biomolecular Screening 12(2): 211-219, 2007

Regulation of gonadotropin release gonadotropin releasing hormone receptors and gonadotrope responsiveness a role for gonadotropin releasing hormone receptor microaggregation. Journal of Cellular Biochemistry 27(1): 13-22, 1985

Development of the high throughput screening assay for identification of agonists of an orphan nuclear receptor. Assay and Drug Development Technologies 4(3): 253-262, 2006

Luteinizing hormone releasing hormone stimulated gonadotropin release from the rainbow trout pituitary gland an in vitro assay for detection of teleost gonadotropin releasing factors. General & Comparative Endocrinology 40(3): 283-290, 1980

Dynamics of gonadotropin releasing hormone stimulated luteinizing hormone release in vitro as related to the behavior of gonadotropin releasing hormone receptor in solution. Clinical Research 28(2): 271A, 1980

Gonadotropin releasing hormone receptor interaction 5. down regulation of pituitary receptors for gonadotropin releasing hormone in ovariectomized ewes by infusion of homologous hormone. Biology of Reproduction 24(5): 1145-1156, 1981

Development of a time-resolved fluorometric assay for the high throughput screening of melanin concentrating hormone receptor antagonists. Journal of Pharmacological and Toxicological Methods 53(3): 242-247, 2005

High-throughput screen for pharmacoperones of the vasopressin type 2 receptor. Journal of Biomolecular Screening 18(8): 930-937, 2013

Pituitary luteinizing hormone responsiveness to gonadotropin releasing hormone in vitro as related to gonadotropin releasing hormone receptor number. American Journal of Physiology 247(5 Part 1): E651-E656, 1984

Pituitary gonadotropin releasing hormone receptor responses to gonadotropin releasing hormone in hypothalamus lesioned rats inhibition of responses by hyperprolactinemia and evidence that testosterone and estradiol modulate gonadotropin secretion at postreceptor sites. Endocrinology 115(3): 1190-1196, 1984