Complex ionic control of [3H]GBR 12783 binding to the dopamine neuronal carrier

Eur J Pharmacol. 1996 Apr 22;301(1-3):195-202. doi: 10.1016/0014-2999(96)00050-7.

Abstract

At 20 degrees C, [3H]GBR 12783, {1-[2-(diphenylmethoxy)ethyl]4-(3-phenyl-2-([1-3H]propenyl)-pip era zine} dissociated from the dopamine neuronal carrier present in rat striatal membranes with a t1/2 value of 27 min. At this temperature, KCI, CaCl2 and MgCl2 increased the binding dissociation, revealing that they recognize a binding site which is not mutually exclusive with that of [3H]GBR 12783. The comparison of the ability of KCl to increase the binding dissociation (by 160% at 30 mM KCl) with its potency as a binding inhibitor (Ki-2.6 +/- 0.3 mM) suggests an involvement of two recognition sites for K+ in binding inhibition, a not mutually exclusive site and another, mutually exclusive, site. Divalent cations mainly inhibited the binding via a mutually exclusive site since 3 mM Ca2+ and 10 mM Mg2+ increased the binding dissociation by 90% at 20 degrees C whereas their Ki values were 0.049 +/- 0.006 and 0.141 +/- 0.035 mM, respectively. Involvement of this mutually exclusive site was also supported by the persistence of the binding inhibition elicited by Ca2+ and Mg2+ at 0 degree C, a temperature at which they reduced the binding dissociation. At 20 degrees C, 100 mM NaCl did not modify [3H]GBR 12783 binding but it antagonized the binding dissociation elicited by inhibitory cations. Ca2+ reduced the off-rate of [3H]GBR 12783 binding at 0 degree C and in the presence of 100 mM Na+. Finally, [3H]GBR 12783-binding dissociation was increased by high 'cytosolic' K+ while 'synaptic' concentrations of Na+, K+, Ca2+, Mg2+ and Cl- were ineffective. A reduction of H2PO4-/HCO3- from 10 to 5 mM and a substitution of 5 mM H2PO4-/HCO3- by 5 mM Cl- increased the binding dissociation, suggesting that an anion-binding site could also regulate the binding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites / drug effects
  • Carrier Proteins / metabolism*
  • Cations / pharmacology
  • Chlorides / pharmacology
  • Dopamine / metabolism*
  • Dopamine Plasma Membrane Transport Proteins
  • Dopamine Uptake Inhibitors / metabolism*
  • In Vitro Techniques
  • Male
  • Membrane Glycoproteins*
  • Membrane Transport Proteins*
  • Nerve Tissue Proteins / metabolism*
  • Neurons / metabolism*
  • Piperazines / metabolism*
  • Protein Binding / drug effects
  • Rats
  • Rats, Sprague-Dawley
  • Temperature

Substances

  • Carrier Proteins
  • Cations
  • Chlorides
  • Dopamine Plasma Membrane Transport Proteins
  • Dopamine Uptake Inhibitors
  • Membrane Glycoproteins
  • Membrane Transport Proteins
  • Nerve Tissue Proteins
  • Piperazines
  • 1-(2-(diphenylmethoxy)ethyl)-4-(3-phenyl-2-propenyl)piperazine
  • Dopamine