Evidence that high- and low-affinity DL-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) binding sites reflect membrane-dependent states of a single receptor

J Neurochem. 1992 Dec;59(6):1997-2004. doi: 10.1111/j.1471-4159.1992.tb10086.x.

Abstract

Binding of DL-alpha-[3H]amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid ([3H]AMPA) to lysed rat brain membranes in the presence of potassium thiocyanate resulted in curvilinear Scatchard plots that could be resolved by regression analysis into a large low-affinity component and a small high-affinity component. Solubilization with Triton X-100 resulted in solubilized and nonsolubilized fractions that were considerably enriched in the high-affinity component and correspondingly reduced in the low-affinity component. It thus appears that solubilization converts low-affinity AMPA receptors into high-affinity receptors. Also, synaptic plasma membranes were found to be greatly enriched in the low-affinity form and deficient in the high-affinity form of the AMPA receptor. These experiments provide evidence for the hypothesis that the high- and low-affinity components of AMPA binding are interconvertible states of the same receptor rather than separate binding sites and that the conversion of these receptors from their native high-affinity state to the low-affinity state occurs on insertion of the receptors into synapses.

Publication types

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

MeSH terms

  • Animals
  • Binding Sites
  • Brain / ultrastructure
  • Ibotenic Acid / analogs & derivatives*
  • Ibotenic Acid / metabolism
  • Male
  • Octoxynol
  • Polyethylene Glycols
  • Protein Binding
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, AMPA
  • Receptors, Glutamate / analysis
  • Receptors, Glutamate / metabolism*
  • Receptors, Glutamate / physiology
  • Solubility
  • Synaptic Membranes / chemistry
  • Synaptic Membranes / metabolism
  • Synaptic Membranes / ultrastructure*
  • alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid

Substances

  • Receptors, AMPA
  • Receptors, Glutamate
  • Ibotenic Acid
  • Polyethylene Glycols
  • alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid
  • Octoxynol