Refolding of misfolded mutant GPCR

Post-translational pharmacoperone action in vitro

Jo Ann Janovick, Shaun P. Brothers, Anda Cornea, Eugene Bush, Mark T. Goulet, Wallace T. Ashton, Daryl R. Sauer, Fortuna Haviv, Jonathan Greer, P. Michael Conn

    Research output: Contribution to journalArticle

    39 Citations (Scopus)

    Abstract

    All reported GnRH receptor mutants (causing human hypogonadotropic hypogonadism) are misfolded proteins that cannot traffic to the plasma membrane. Pharmacoperones correct misfolding and rescue mutants, routing them to the plasma membrane where they regain function. Because pharmacoperones are often peptidomimetic antagonists, these must be removed for receptor function after rescue; in vivo this necessitates pulsatile pharmacoperone administration. As an antecedent to in vivo studies, we determined whether pharmacoperones need to be present at the time of synthesis or whether previously misfolded proteins could be refolded and rescued. Accordingly, we blocked either protein synthesis or intra-cellular transport. Biochemical and morphological studies using 12 mutants and 10 pharmacoperones representing three different chemical classes show that previously synthesized mutant proteins, retained by the quality control system (QCS), are rescued by pharmacoperones, showing that pharmacoperone administration in vivo likely need not consider whether the target protein is being synthesized at the time of drug administration.

    Original languageEnglish (US)
    Pages (from-to)77-85
    Number of pages9
    JournalMolecular and Cellular Endocrinology
    Volume272
    Issue number1-2
    DOIs
    StatePublished - Jun 30 2007

    Fingerprint

    Cell membranes
    Proteins
    Cell Membrane
    Peptidomimetics
    LHRH Receptors
    Regain
    Hypogonadism
    Mutant Proteins
    Quality Control
    Quality control
    Control systems
    In Vitro Techniques
    Pharmaceutical Preparations

    Keywords

    • G protein coupled receptor
    • Hypogonadotropic hypogonadism
    • Protein folding
    • Protein trafficking

    ASJC Scopus subject areas

    • Endocrinology
    • Endocrinology, Diabetes and Metabolism

    Cite this

    Janovick, J. A., Brothers, S. P., Cornea, A., Bush, E., Goulet, M. T., Ashton, W. T., ... Michael Conn, P. (2007). Refolding of misfolded mutant GPCR: Post-translational pharmacoperone action in vitro. Molecular and Cellular Endocrinology, 272(1-2), 77-85. https://doi.org/10.1016/j.mce.2007.04.012

    Refolding of misfolded mutant GPCR : Post-translational pharmacoperone action in vitro. / Janovick, Jo Ann; Brothers, Shaun P.; Cornea, Anda; Bush, Eugene; Goulet, Mark T.; Ashton, Wallace T.; Sauer, Daryl R.; Haviv, Fortuna; Greer, Jonathan; Michael Conn, P.

    In: Molecular and Cellular Endocrinology, Vol. 272, No. 1-2, 30.06.2007, p. 77-85.

    Research output: Contribution to journalArticle

    Janovick, JA, Brothers, SP, Cornea, A, Bush, E, Goulet, MT, Ashton, WT, Sauer, DR, Haviv, F, Greer, J & Michael Conn, P 2007, 'Refolding of misfolded mutant GPCR: Post-translational pharmacoperone action in vitro', Molecular and Cellular Endocrinology, vol. 272, no. 1-2, pp. 77-85. https://doi.org/10.1016/j.mce.2007.04.012
    Janovick, Jo Ann ; Brothers, Shaun P. ; Cornea, Anda ; Bush, Eugene ; Goulet, Mark T. ; Ashton, Wallace T. ; Sauer, Daryl R. ; Haviv, Fortuna ; Greer, Jonathan ; Michael Conn, P. / Refolding of misfolded mutant GPCR : Post-translational pharmacoperone action in vitro. In: Molecular and Cellular Endocrinology. 2007 ; Vol. 272, No. 1-2. pp. 77-85.
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