In conclusion, a novel recognition motif between anionic pillararene (2C-WP5A) and azobenzene derivative G has been developed, and both of the trans- and cis- G have the similar affinity with 2C-WP5A. The complexation process was well studied and confirmed by 1H NMR, 2D ROESY, and ITC experiment. The recognition motif would greatly expand the application in the area of construction of stimuli-responsive supramolecular assembly based on anionic pillararene. Furthermore, we utilized this novel recognition motif to construct supramolecular assembly nanoparticles which could reversibly switch the size from ~800 nm to ~250 nm by photo/thermal stimulation, presenting the turbid-to-clear switch of the solution state with excellent reversibility. To the best of our knowledge, this is the first example of supramolecular aggregate based anionic pillararene and azobenzene derivative. The high affinity of anionic pillararene with azobenzene derivative is expected to have great potential applications in further fabrication of more sophisticated stimuli-responsive supramolecular system.