A major focus of current research on the nicotinic acetylcholine receptor (AChR) has been to understand the molecular mechanism of ion channel inhibition. In particular, we put special emphasis on the description of the localization of the agonist self-inhibitory binding site. Binding of agonist in the millimolar concentration range to this particular site produces inhibition of the ion flux activity previously elicited by the same agonist at micromolar concentrations. Due to the similitude in the pharmacological and electrophysiological behavior in inhibiting the ion channel of both high agonist concentrations and noncompetitive antagonists, we first describe the localization of noncompetitive inhibitor binding sites on the AChR. There is a great body of experimental evidence for the existence and location of luminal high-affinity noncompetitive inhibitor binding sites. In this regard, the most simple mechanism to describe the action of noncompetitive inhibitors which bind to luminal sites and, by its semblance, the agonist self-inhibition itself, is based on the assumption that these compounds enter the open channel, bind to different rings within the M2 transmembrane domain of the receptor, and block cation flux by occluding the receptor pore. However, the existence of high-affinity nonluminal noncompetitive inhibitor binding sites is not consistent with the open-channel-blocking mechanism. Instead, the presence of the quinacrine locus at the lipid-protein (alpha M1) interface approximately 7 A from the lipid-water interface and the ethidium domain located approximately 46 A from the membrane surface in the wall of the vestibule open the possibility for the regulation of cation permeation by an allosteric process. Additionally, the observed (at least partially) overlapping between the quinacrine and the agonist self-inhibitory binding site also suggests an allosteric process for agonist self-inhibition. For this alternative mechanism, cholinergic agonist molecules first need to be partitioned into (or to be adsorbed onto) the lipid membrane to further interact with its binding site located at the lipid-protein interface.