HSP90 Regulation of P2X7 Receptor Function Requires an Intact Cytoplasmic C-Terminus

Mol Pharmacol. 2016 Aug;90(2):116-26. doi: 10.1124/mol.115.102988. Epub 2016 Jun 14.

Abstract

P2X7 receptors (P2X7Rs) are ATP-gated ion channels that display the unusual property of current facilitation during long applications of agonists. Here we show that facilitation disappears in chimeric P2X7Rs containing the C-terminus of the P2X2 receptor (P2X2R), and in a truncated P2X7R missing the cysteine-rich domain of the C-terminus. The chimeric and truncated receptors also show an apparent decreased permeability to N-methyl-d-glucamine(+) (NMDG(+)). The effects of genetic modification of the C-terminus on NMDG(+) permeability were mimicked by preapplication of the HSP90 antagonist geldanamycin to the wild-type receptor. Further, the geldanamycin decreased the shift in the reversal potential of the ATP-gated current measured under bi-ionic NMDG(+)/Na(+) condition without affecting the ability of the long application of agonist to facilitate current amplitude. Taken together, the results suggest that HSP90 may be essential for stabilization and function of P2X7Rs through an action on the cysteine-rich domain of the cytoplasmic the C-terminus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / pharmacology
  • Animals
  • Benzoquinones / pharmacology
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism
  • Cell Membrane Permeability / drug effects
  • Cytoplasm / metabolism*
  • HEK293 Cells
  • HSP90 Heat-Shock Proteins / metabolism*
  • Humans
  • Ion Channel Gating / drug effects
  • Lactams, Macrocyclic / pharmacology
  • Meglumine / metabolism
  • Mutant Proteins / metabolism
  • Protein Binding / drug effects
  • Protein Domains
  • Rats
  • Receptors, Purinergic P2X7 / chemistry*
  • Receptors, Purinergic P2X7 / metabolism*
  • Recombinant Proteins / metabolism
  • Structure-Activity Relationship

Substances

  • Benzoquinones
  • HSP90 Heat-Shock Proteins
  • Lactams, Macrocyclic
  • Mutant Proteins
  • Receptors, Purinergic P2X7
  • Recombinant Proteins
  • Meglumine
  • Adenosine Triphosphate
  • geldanamycin