Thermodynamic analysis of agonist and antagonist binding to membrane-bound and solubilized A1 adenosine receptors

J Pharmacol Exp Ther. 1993 Sep;266(3):1463-74.

Abstract

The thermodynamic properties of the agonist [Adenine-2,8-3H, ethyl-2(3)-H]-N6-phenylisopropyladenosine ([3H]R-PIA) and the antagonist 8-Cyclopentyl-1,3-[3H]dipropylxanthine ([3H]DPCPX) binding to membrane-bound and 3-3[-(choloamidopropyl)-dimethylammoniol-1-propanesulfonate/ digitonin- solubilized A1 adenosine receptors from pig brain cortex were evaluated. Rate constants for [3H]R-PIA and [3H]DPCPX association (k+1) and dissociation (k-1) processes to this receptor subtype were measured from association-dissociation experiments at six different temperatures. The values for equilibrium association constant (KA = 1/KD) were derived from rate constant values (k+1/k-1). The antagonist binding to membrane-bound receptors, the agonist binding to fast kinetic component membrane-bound receptors and the agonist binding to soluble receptors showed a linear temperature-dependence of the standard free-energy change. The first two processes are enthalpy- and entropy-driven, and the third process is enthalpy-driven with entropy working against it. On the other hand, a curvilinear temperature-dependence appears in the agonist binding to slow kinetic component membrane-bound receptors and in the antagonist binding to soluble receptors, but analyzing the semireactions (association-dissociation) involved in each case reveals that the thermodynamic behavior is very different. The thermodynamic similarities and differences are discussed in terms of receptor--G protein interaction.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Kinetics
  • Membranes / metabolism
  • Membranes / ultrastructure
  • Phenylisopropyladenosine / chemistry*
  • Phenylisopropyladenosine / metabolism*
  • Postural Balance
  • Receptors, Purinergic / chemistry
  • Receptors, Purinergic / metabolism*
  • Receptors, Purinergic / physiology
  • Solutions
  • Swine
  • Temperature
  • Thermodynamics
  • Tritium
  • Xanthines / chemistry*
  • Xanthines / metabolism*

Substances

  • Receptors, Purinergic
  • Solutions
  • Xanthines
  • Tritium
  • Phenylisopropyladenosine
  • 1,3-dipropyl-8-cyclopentylxanthine