Functional rescue of Kallmann syndrome-associated prokineticin receptor 2 (PKR2) mutants deficient in trafficking

J Biol Chem. 2014 May 30;289(22):15518-26. doi: 10.1074/jbc.M114.556381. Epub 2014 Apr 21.

Abstract

Mutations in the G protein-coupled prokineticin receptor 2 (PKR2) are known to cause Kallmann syndrome and idiopathic hypogonadotropic hypogonadism manifesting with delayed puberty and infertility. Some of the mutant receptors are not routed to the cell surface; instead, they are trapped in the cellular secretory pathway. The cell-permeant agonists/antagonists have been used to rescue some membrane receptors that are not targeted onto the cell membrane. Here, we chose three disease-associated mutations (W178S, G234D, and P290S), which all resulted in retention of PKR2 intracellularly. We show that a small molecule PKR2 antagonist (A457) dramatically increased cell surface expression and rescued the function of P290S PKR2, but had no effect on W178S and G234D PKR2. Furthermore, we also tested chemical chaperone glycerol on the cell surface expression and function of PKR2 mutants. Treatment with 10% glycerol significantly increased the cell surface expression and signaling of P290S and W178S PKR2. These data demonstrate that some Kallmann syndrome-associated, intracellularly retained mutant PKR2 receptors can be functionally rescued, suggesting a potential treatment strategy for patients bearing such mutations.

Keywords: G Protein-coupled Receptor (GPCR); Kallmann Syndrome; Molecular Chaperone; PKR2; Protein Misfolding; Signal Transduction; Trafficking.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cricetulus
  • Cryoprotective Agents / pharmacology
  • Glycerol / pharmacology
  • HEK293 Cells
  • Heterocyclic Compounds, 4 or More Rings / chemical synthesis
  • Heterocyclic Compounds, 4 or More Rings / pharmacology*
  • Humans
  • Kallmann Syndrome / drug therapy
  • Kallmann Syndrome / genetics*
  • Kallmann Syndrome / metabolism*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Point Mutation
  • Protein Transport / genetics
  • Proteostasis Deficiencies / genetics
  • Proteostasis Deficiencies / metabolism
  • Receptors, G-Protein-Coupled / antagonists & inhibitors
  • Receptors, G-Protein-Coupled / genetics*
  • Receptors, G-Protein-Coupled / metabolism*
  • Receptors, Peptide / antagonists & inhibitors
  • Receptors, Peptide / genetics*
  • Receptors, Peptide / metabolism*
  • Signal Transduction / drug effects
  • Signal Transduction / genetics

Substances

  • A457 compound
  • Cryoprotective Agents
  • Heterocyclic Compounds, 4 or More Rings
  • Membrane Proteins
  • PROKR2 protein, human
  • Receptors, G-Protein-Coupled
  • Receptors, Peptide
  • Glycerol