Fig. 5: Characterization of artificial molecular signaling system.

a Schematic illustration of AMSsys working principle. Initially, chol-c-DNGK with biomimetic GPMV effectively shields the extracellular zinc ions. Driven by ATP, however, chol-c-DNGK switches to the open state, allowing ions to freely flow into the chamber by diffusion and trigger the signaling network. Eventually, the feedback pathway mediates chol-o-DNGK to switch back to the original closed state. Simultaneously, reactions of confined signaling network have been accomplished. b CLSM analysis of AMSsys in the above different states. Panel 1: without ATP and ions. Confined signaling network stays inactivated: Cy5-labeled (633 channel) blocker is free in the chamber, where the 488 signal saw only minor increase. Panel 2: Addition of ATP drives the opening of chol-c-DNGK, while the signaling network continues to remain inactive by the absence of ion influx triggering. Panel 3: Addition of only ions (no ATP) cannot trigger the confined reactive network and chol-DNGK remains closed. Panel 4: Only in the presence of both ATP and ion influx, can all designed reactions be completed: chol-c-DNGK switches to chol-o-DNGK, followed by diffusion of ions through the mimetic nanochannel, triggering, in turn, the confined signaling network. After completion of the confined reaction cascade, the fluorescent signal from SG-I is revealed (light green spot) and Cy5-labeled blocker plugs the nanogatekeeper (red circle). Plus (“+”) mark denotes that the corresponding trigger (ATP or/and ions) was present. Scale bar, 5 μM. c Fluorescence intensity of the white dashed line section of each group in b. Groups I, II, III, and IV, respectively, represent the independent groups (from panel 1 to panel 4) in b from top to bottom. The green pattern shows the SG-I fluorescence channel, while the red pattern shows the Cy5 fluorescence channel. Source data are provided in the Source Data file.