Defining affinity with the GABA(A) receptor

Mathew V. Jones, Yoshinori Sahara, Jeffrey A. Dzubay, Gary Westbrook

Research output: Contribution to journalArticle

132 Citations (Scopus)

Abstract

At nicotinic and glutamatergic synapses, the duration of the postsynaptic response depends on the affinity of the receptor for transmitter (Colquhoun et al., 1977; Pan et al., 1993). Affinity is often thought to be determined by the ligand unbinding rate, whereas the binding rate is assumed to be diffusion-limited. In this view, the receptor selects for those ligands that form a stable complex on binding, but binding is uniformly fast and does not itself affect selectivity. We tested these assumptions for the GABA(A) receptor by dissecting the contributions of microscopic binding and unbinding kinetics for agonists of equal efficacy but of widely differing affinities. Agonist pulses applied to outside-out patches of cultured rat hippocampal neurons revealed that agonist unbinding rates could not account for affinity if diffusion-limited binding was assumed. However, direct measurement of the instantaneous competition between agonists and a competitive antagonist revealed that binding rates were orders of magnitude slower than expected for free diffusion, being more steeply correlated with affinity than were the unbinding rates. The deviation from diffusion-limited binding indicates that a ligand-specific energy barrier between the unbound and bound states determines GABA(A) receptor selectivity. This barrier and our kinetic observations can be quantitatively modeled by requiring the participation of movable elements within a flexible GABA binding site.

Original languageEnglish (US)
Pages (from-to)8590-8604
Number of pages15
JournalJournal of Neuroscience
Volume18
Issue number21
StatePublished - Nov 1 1998

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GABA-A Receptors
Ligands
Synapses
gamma-Aminobutyric Acid
Binding Sites
Neurons

Keywords

  • Kinetics
  • Ligand-gated channels
  • Molecular modeling
  • Selectivity
  • Synapse
  • Thermodynamic

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Jones, M. V., Sahara, Y., Dzubay, J. A., & Westbrook, G. (1998). Defining affinity with the GABA(A) receptor. Journal of Neuroscience, 18(21), 8590-8604.

Defining affinity with the GABA(A) receptor. / Jones, Mathew V.; Sahara, Yoshinori; Dzubay, Jeffrey A.; Westbrook, Gary.

In: Journal of Neuroscience, Vol. 18, No. 21, 01.11.1998, p. 8590-8604.

Research output: Contribution to journalArticle

Jones, MV, Sahara, Y, Dzubay, JA & Westbrook, G 1998, 'Defining affinity with the GABA(A) receptor', Journal of Neuroscience, vol. 18, no. 21, pp. 8590-8604.
Jones MV, Sahara Y, Dzubay JA, Westbrook G. Defining affinity with the GABA(A) receptor. Journal of Neuroscience. 1998 Nov 1;18(21):8590-8604.
Jones, Mathew V. ; Sahara, Yoshinori ; Dzubay, Jeffrey A. ; Westbrook, Gary. / Defining affinity with the GABA(A) receptor. In: Journal of Neuroscience. 1998 ; Vol. 18, No. 21. pp. 8590-8604.
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