Fig. 2: Comparison of the stepping behavior for constructs labeled at different amino acids.
a Exemplary on-axis position-time traces of construct T324C (inset) labeled at the center-right of the head domain recorded at 50 µM (purple) and 5 mM (green) ATP concentration. The raw position data (semi-transparent lines) are overlaid with a step-fit (solid lines). The zoom-in highlights 16 nm regular steps and unequally-sized substeps. b Population-normalized histograms of measured step sizes recorded at the two ATP concentrations showing step sizes of 16 nm and substeps between 6 nm and 11 nm (N50µM = 1625, N5mM = 1592). c Comparison of the on-axis stepping behavior between construct T324C and K28C (constructs depicted on the left). The sequence of step sizes is displayed by a bivariate density scatter plot (middle). The step sizes recorded after a regular step (16 nm) are underlaid in dark blue, those before a regular step in light blue. The projections along the short axis of these boxes are shown as plots (right) showing similar substep sizes for construct K28C and varying substep sizes for construct T324C. The total number of steps for construct T324C and K28C are given by NT324C = 3301 and NK28C = 5519, respectively. d Comparison of the off-axis stepping behavior. Bivariate density scatter plots of the off-axis displacement for all subsequent bound to unbound and unbound to bound substeps together with the corresponding ellipses from the eigenvalues and eigenvectors of the covariance matrices (NT324C = 590, NK28C = 1055). Data for construct T324C is taken from 216 traces of 12 biological replicates (for statistics on construct K28C refer to Fig. 1).
