1. In diverse tissues, acetylcholinesterase appears to play a critical role in the functional state of cells completely dependent of cholinergic transmission. However, very little is known about the mechanisms and actual molecular structures mediating the fundamental interactions between this protein and the cellular membrane. 2. In this study, peritoneal macrophages were used as a model system to study the possible interaction between acetylcholinesterase, acting in a non-cholinergic capacity, and the cellular membrane. 3. When acetylcholinesterase was incubated with macrophages harvested from rat peritoneum, the rate of oxygen consumption was increased in a concentration-dependent manner that was independent of mitochondrial block with sodium cyanide. Furthermore, heat inactivation of enzymatic activity or application of BW 284C51 at a concentration which totally blocks catalytic activity did not eliminate the effect. 4. In contrast, incubation with bovine serum albumin or butyrylcholinesterase actually retarded oxygen consumption. 5. The effect of acetylcholinesterase depended on the presence of divalent cations and was inhibited by mannan and D-mannose, but not D-galactose. It is concluded that acetylcholinesterase can induce a "respiratory burst" in macrophages independent of its conventional catalytic site but involving either the mannose receptor of the monocyte-derived macrophage or a possible sugar binding site on acetylcholinesterase itself.