A2A adenosine receptors were examined in bovine striatal membranes following exposure to tetranitromethane (TNM) which modifies tyrosine and cysteine residues. TNM (0.05-0.5 mM) treatment caused an irreversible, concentration-dependent decrease in the binding activity of the selective A2A agonist [3H]CGS 21680. Protection studies showed that TNM inactivation could be prevented by the adenosine receptor agonist 5'-N-ethylcarboxamidoadenosine (NECA) and by the antagonist xanthine amine congener (XAC), suggesting that TNM modified residues at the ligand-binding sites. Scatchard analysis of the binding data showed that 0.15 mM TNM decreased the [3H]CGS 21680 Bmax value from 447 +/- 39 to 273 +/- 21 fmol/mg of proteins without any significant change in the Kd values (13.5 +/- 1.4 and 14.7 +/- 1.5 for control and treated membranes, respectively). We carried out a series of successive chemical modifications with the reducing agent dithiothreitol (DTT), which indicated that the residues modified by TNM, under our experimental conditions, are tyrosine residues and not cysteine residues.