N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine inhibits ligand binding to certain G protein-coupled receptors.

N,N,N',N'-Tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) is used widely in biological systems to chelate certain heavy metals, particularly Zn2+. Here we show that TPEN inhibits ligand binding to certain G protein-coupled receptors and is an antagonist at muscarinic receptors. In intact human neuroblastoma SH-SY5Y cells, the binding of the muscarinic receptor ligand [N-methyl-3H]scopolamine methyl chloride was inhibited by TPEN (Ki approximately 26 microM), as was muscarinic receptor agonist-induced inositol 1,4,5-trisphosphate formation (Ki approximately 26 microM). This antagonism was not due to metal ion chelation, indicating that it resulted from a direct interaction of TPEN with muscarinic receptors. Examination of the effects of TPEN on other receptors in SH-SY5Y cell membrane preparations showed that the binding of the nonpeptide opioid receptor ligand [15,16-3H]diprenorphine was strongly inhibited, whereas binding of [125I]vasoactive intestinal polypeptide was not. This pattern of selectivity was also seen in AR4-2J rat pancreatoma cell membranes, in which TPEN inhibited ligand binding to muscarinic receptors, but not that to cholecystokinin receptors. In conclusion, these data show that TPEN inhibits ligand binding to certain G protein-coupled receptors and exhibits selectivity towards those receptors whose transmembrane helices form the predominant site for ligand interaction. TPEN may have widespread antagonistic activity towards G protein-coupled receptors of this kind.