Membrane vesicles (ghosts) containing ascorbic acid were prepared from bovine chromaffin granules. When ferricyanide or ferricytochrome c were added to the external medium, a membrane potential (interior positive) developed across the ghost membrane. This membrane potential could not be elicited from ascorbate-free ghosts or by ferrocyanide added instead of ferricyanide. These results indicate that the chromaffin-granule membrane has a transmembrane electron carrier with a midpoint potential between that of ascorbate (+85 mV) and that of cytochrome c (+255 mV). The most likely candidate is cytochrome b-561 (+140 mV).