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

40 Scopus citations

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
Externally publishedYes

Keywords

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

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

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    Janovick, J. A., Brothers, S. P., Cornea, A., Bush, E., Goulet, M. T., Ashton, W. T., Sauer, D. R., Haviv, F., Greer, J., & 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