PIP2 regulates psychostimulant behaviors through its interaction with a membrane protein

Nat Chem Biol. 2014 Jul;10(7):582-589. doi: 10.1038/nchembio.1545. Epub 2014 Jun 1.

Abstract

Phosphatidylinositol (4,5)-bisphosphate (PIP2) regulates the function of ion channels and transporters. Here, we demonstrate that PIP2 directly binds the human dopamine (DA) transporter (hDAT), a key regulator of DA homeostasis and a target of the psychostimulant amphetamine (AMPH). This binding occurs through electrostatic interactions with positively charged hDAT N-terminal residues and is shown to facilitate AMPH-induced, DAT-mediated DA efflux and the psychomotor properties of AMPH. Substitution of these residues with uncharged amino acids reduces hDAT-PIP2 interactions and AMPH-induced DA efflux without altering the hDAT physiological function of DA uptake. We evaluated the significance of this interaction in vivo using locomotion as a behavioral assay in Drosophila melanogaster. Expression of mutated hDAT with reduced PIP2 interaction in Drosophila DA neurons impairs AMPH-induced locomotion without altering basal locomotion. We present what is to our knowledge the first demonstration of how PIP2 interactions with a membrane protein can regulate the behaviors of complex organisms.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Substitution
  • Amphetamine / pharmacology*
  • Animals
  • Behavior, Animal / drug effects*
  • Cell Membrane / drug effects
  • Central Nervous System Stimulants / pharmacology*
  • Dopamine / metabolism
  • Dopamine Plasma Membrane Transport Proteins / chemistry
  • Dopamine Plasma Membrane Transport Proteins / genetics
  • Dopamine Plasma Membrane Transport Proteins / metabolism*
  • Drosophila melanogaster / drug effects*
  • Drosophila melanogaster / physiology
  • Gene Expression
  • Humans
  • Locomotion / drug effects
  • Models, Molecular
  • Neurons / drug effects
  • Neurons / metabolism
  • Phosphatidylinositol 4,5-Diphosphate / metabolism*
  • Phosphatidylinositol 4,5-Diphosphate / pharmacology
  • Protein Structure, Tertiary
  • Transgenes

Substances

  • Central Nervous System Stimulants
  • Dopamine Plasma Membrane Transport Proteins
  • Phosphatidylinositol 4,5-Diphosphate
  • Amphetamine
  • Dopamine