Journal article
Rescue of expression and signaling of human luteinizing hormone G protein-coupled receptor mutants with an allosterically binding small-molecule agonist
Proceedings of the National Academy of Sciences - PNAS, Vol.108(17), pp.7172-7176
04/26/2011
DOI: 10.1073/pnas.1015723108
PMCID: PMC3084114
PMID: 21482767
Abstract
Naturally occurring mutations of G protein-coupled receptors (GPCRs) causing misfolding and failure to traffic to the cell surface can result in disease states. Some small-molecule orthosteric ligands can rescue such misfolded receptors, presumably by facilitating their correct folding and shuttling to the plasma membrane. Here we show that a cell-permeant, allosterically binding small-molecule agonist (Org 42599) rescues the folding and cell surface expression, and therefore target cell signaling, of mutant human luteinizing hormone (LH) receptors (A593P and S616Y) that cause Leydig cell hypoplasia in man. Both mutant receptors were retained in the cytoplasm whereas WT receptor localized at the cell membrane, and binding of LH to cells expressing the mutant receptors was markedly lower than to those expressing the WT receptor. Incubation with Org 42599 increased mutant receptor expression, cell surface localization, and the proportion of mutant receptor in the mature glycosylated form. Importantly, although LH stimulated little (S616Y) or no (A593P) activation of cells expressing mutant receptors, incubation of cells with Org 42599 facilitated rescue of expression and stimulation by the native ligand, LH. Although Org 42599 could activate these receptors, it could not displace
125
I-labeled human LH binding to the WT receptor, indicating that it acts in an allosteric manner. Here we demonstrate a small-molecule GPCR allosteric agonist that functionally rescues intracellularly retained mutant LH receptors by facilitating their cell surface expression. This approach may have application for treatment of infertile patients bearing such mutations and, more broadly, for other misfolded GPCR mutants resulting in human pathologic processes.
Details
- Title: Subtitle
- Rescue of expression and signaling of human luteinizing hormone G protein-coupled receptor mutants with an allosterically binding small-molecule agonist
- Creators
- Claire L Newton - Centre for Integrative Physiology, School of Biomedical SciencesAdele M Whay - , Glasgow G81 2LGCraig A McArdle - Laboratories for Integrative Neuroscience and Endocrinology (LINE), School of Clinical SciencesMeilin Zhang - Department of Molecular Biophysics and Physiology, The Roy J. and Lucille A. Carver College of MedicineChris J van Koppen - Molecular Pharmacology and Drug Metabolism and Pharmacokinetics, Women's Health DepartmentRuud van de Lagemaat - Molecular Pharmacology and Drug Metabolism and Pharmacokinetics, Women's Health DepartmentDeborah L Segaloff - Department of Molecular Biophysics and Physiology, The Roy J. and Lucille A. Carver College of MedicineRobert P Millar - Centre for Integrative Physiology, School of Biomedical Sciences
- Resource Type
- Journal article
- Publication Details
- Proceedings of the National Academy of Sciences - PNAS, Vol.108(17), pp.7172-7176
- DOI
- 10.1073/pnas.1015723108
- PMID
- 21482767
- PMCID
- PMC3084114
- NLM abbreviation
- Proc Natl Acad Sci U S A
- ISSN
- 0027-8424
- eISSN
- 1091-6490
- Publisher
- National Academy of Sciences
- Language
- English
- Date published
- 04/26/2011
- Academic Unit
- Molecular Physiology and Biophysics; Obstetrics and Gynecology
- Record Identifier
- 9984083893802771
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