In this study, the charge selectivity of staphylococcal ␣-hemolysin (␣HL), a bacterial pore-forming toxin, is manipulated by using cyclodextrins as noncovalent molecular adapters. Anionselective versions of ␣HL, including the wild-type pore and various mutants, become more anion selective when -cyclodextrin (CD) is lodged within the channel lumen. By contrast, the negatively charged adapter, hepta-6-sulfato--cyclodextrin (s7CD), produces cation selectivity. The cyclodextrin adapters have similar effects when placed in cation-selective mutant ␣HL pores. Most probably, hydrated Cl ؊ ions partition into the central cavity of CD more readily than K ؉ ions, whereas s7CD introduces a charged ring near the midpoint of the channel lumen and confers cation selectivity through electrostatic interactions. The molecular adapters generate permeability ratios (P K؉/PCl؊) over a 200-fold range and should be useful in the de novo design of membrane channels both for basic studies of ion permeation and for applications in biotechnology.