Binding of radiolabeled antagonist to platelet-activating-factor receptor in human lung membranes: shifting of agonist binding affinity states by cations and guanine nucleotide.

Binding of the radiolabeled platelet-activating-factor (PAF) receptor antagonist RP52770, [( 3H]-N-(3-chlorophenyl)-3-(3-pyridinyl)-1H, 3H-pyrrolo- [1,2-c]thiazole-7-carboxamide) to receptors in human lung membranes was time- dependent, protein-dependent, reversible and saturable. The dissociation constant and maximal binding density were 14 +/- 2 nM and 2.1 +/- 0.6 pmol/mg protein, respectively. [3H]-RP52770 binding to the PAF receptor was competitively displaced by PAF and receptor antagonists. The rank order of the binding affinities were PAF greater than RP52770 (+) greater than RP52770 (-) greater than CV3988, equivalent to the PAF receptor specificities determined from functional studies. Binding of PAF to [3H]-RP52770 labeled receptors was regulated by sodium, guanylylimido- diphosphate (GppNHp) and divalent cations. In the presence of EDTA, Na+ and GppNHp, in combination, binding of PAF to the receptor was maximally shifted to the right. These results clearly demonstrate that cations and guanine nucleotide can regulate the affinity states of the PAF receptor in human lung membranes.