Figure 6.

Single molecule tracking analyses of the MTase and REase, expressed at very low levels as mVenus fusions. (A) Representative tracks (in yellow) of enzyme molecules, in cells that are outlined by white ovals. (B) Projection of all tracks into a standardized cell of 3×1 μm size. Upper panels are heat maps, where darker shading indicates higher presence of molecule tracks. Lower panels are confinement maps, with tracks: (i) moving within a radius of 120 nm for at least five steps are shown in red, (ii) tracks moving freely in blue and (iii) tracks containing both confined and free motion in green (‘transitions’). (C) Jump distance diagrams showing the cumulative probability distribution of squared displacement analyses (SQD) to estimate the diffusion constants (D) and relative fractions of up to three diffusive states. Upper panels show data fitted with two Rayleigh distributions, lower panels with three distributions. Inset show deviation (blue line) of experimental data from modelled data (indicated by dashed red line). (D) Summary of data obtained from SQD analyses, diffusion constants D_1–3 correspond to populations Pop_1–3.