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. 2016 Sep 20;7:12789. doi: 10.1038/ncomms12789

Figure 2. MloK1 dynamically changes protrusion height during HS-AFM imaging.

Figure 2

(a) Representative HS-AFM movie frames of the same MloK1-containing membrane. Arrows 1 and 2 highlight two individual molecules continuously blinking between two heights. On the right are the corresponding height profiles along the centre axis of the outlines in each frame on the left. During blinking the CNBDs undergo a height change of ∼1 nm. Scale bar: 20 nm. Full colour scale: 2.5 nm. (b) Cartoon of the blinking process: unliganded CNBDs alternate between down- and up-states. The kymograph (lower) plots the height evolution over 3 min for the eight MloK1 molecules highlighted in a. Colour display is identical. (c) Height histograms of the non-blinking (grey) and blinking (red) molecule from the kymograph in b (arrows). While the non-blinking channels show a one-peak distribution, the blinking channels display a broad height histogram. (d) High-resolution HS-AFM image frames showing an example of individual blinking subunits (arrows) within an MloK1 tetramer. Scale bar: 10 nm. Full colour scale: 2 nm. Buffer contains 10 nM cAMP for a–c, and 100 μM cAMP for d.