Rescue of misrouted GnRHR mutants reveals its constitutive activity

Jo Ann Janovick, Irina D. Pogozheva, Henry I. Mosberg, Anda Cornea, P. Michael Conn

    Research output: Contribution to journalArticle

    5 Scopus citations

    Abstract

    G protein-coupled receptors (GPCR) play central roles in almost all physiological functions, and mutations in GPCR are responsible for over 30 hereditary diseases associated with loss or gain of receptor function. Gain of function mutants are frequently described as having constitutive activity (CA), that is, they activate effectors in the absence of agonist occupancy. Although many GPCR have mutants with CA, the GnRH receptor (GnRHR) was not, until 2010, associated with any CA mutants. The explanation for the failure to observe CA appears to be that the quality control system of the cell recognizes CA mutants of GnRHR as misfolded and retains them in the endo-plasmic reticulum. In the present study, we identified several human (h)GnRHR mutants with substitutions in transmembrane helix 6 (F272K, F272Q, Y284F, C279A, and C279S) that demonstrate varying levels of CA after being rescued by pharmacoperones from different chemical classes and/or deletionofresidue K191,a modification that increases traffickingtothe plasma membrane. The movement of the mutants from the endoplasmic reticulum (unrescued) to the plasma membrane (after rescue) is supported by confocal microscopy. Judging from the receptor-stimulated inositol phosphate production, mutants F272K and F272Q, after rescue, display the largest level of CA, an amount that is comparable with agonist-stimulated activation. Because mutations in other GPCR are, like the hGnRHR, scrutinized by the quality control system, this general approach may reveal CA in receptor mutants from other systems. A computer model of the hGnRHR and these mutants was used to evaluate the conformation associated with CA.

    Original languageEnglish (US)
    Pages (from-to)1179-1188
    Number of pages10
    JournalMolecular Endocrinology
    Volume26
    Issue number7
    DOIs
    StatePublished - Jul 2012

    ASJC Scopus subject areas

    • Molecular Biology
    • Endocrinology

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