7 alpha-Bis (beta-chloroethyl)amino-methyl-6,14-endoethenotetrahydrooripavine (alpha-CAM) was found to bind to opioid receptors irreversibly and react directly with sulfhydryl (SH) groups in P2 preparations of rat brain. The P2 preparations were pretreated as follows: protection of the SH groups at the opioid receptor binding sites by morphine or etorphine, and inactivation of the SH groups outside the binding sites by N-ethylmaleimide (NEM), followed by removal of the morphine or etorphine by washing. alpha-CAM was still able to bind the pretreated P2 preparations in an irreversible manner. The results indicate that the formation of covalent bonds between alpha-CAM and the SH groups of opioid receptor binding sites is possibly one of the biochemical mechanisms of the irreversible action of alpha-CAM.