The open-close state of the ion channels in a living system are regulated by multiple stimuli such as ligand,pH,potential and light.Functionalizing natural channels by using synthetic chemistry would provide biological nanopores with novel properties and applications.Here we use para-sulfonato-calix[4]arene-based host-guest supramolecular system to develop artificial gating mechanisms aiming at regulating wild-type α-HL commanded by both ligand and light stimuli.Using the gating p ope ty of α-hemolysin,we studied the host-guest interactions between para-sulfonato-calix[4]arene and 4,4-dipyridinium-azobenzene at the single-molecule level.Subsequently,we have extended the application of this gating system to the real-time study of light-induced molecule shuttle based on para-sulfonato-calix[4]arene and 4,4-dipyridinium-azobenzene at single-molecule level.These experiments provide a more efficient method to develop a general tool to analyze the individual motions of supramolecul systems by using comme ci lly il ble α-HL nanopores.