Skip to main content
. 2019 Sep 13;294(45):17017–17030. doi: 10.1074/jbc.RA119.008947

Figure 2.

Figure 2.

Dwell time analysis of main pause and subpause. A, [ATP] dependence of rotation velocity of WT EhV1. The red open circles and small filled circles show the average velocity and velocities of individual molecules, respectively. Red line, fit with the Michaelis–Menten equation: V = VmaxATP × [ATP]/(KmATP + [ATP]). The VmaxATP and KmATP were 117 ± 3 rps (fitted parameter ± fitting error) and 43 ± 6 μm, respectively. The konATP was estimated from 3 × VmaxATP/KmATP as 8.2 × 106 m−1 s−1. B, distributions of duration of the main pause. For 100 nm and 1 μm ATP, the distributions were fitted with single-exponential decay functions: constant × exp(−t/τ). The time constants were obtained as 972 ± 41 and 95.7 ± 3.7 ms for 100 nm and 1 μm, respectively. From 10 μm to 30 mm ATP, the distributions were fitted with double-exponential decay functions assuming two consecutive first-order reactions: constant × (exp(−t/τ) − exp(−t/τ′)). The time constants were obtained as 11.9 ± 0.8 and 1.1 ± 0.2 ms for 10 μm, 1.4 ± 0.9 and 1.0 ± 0.9 ms for 100 μm, 0.7 ± 0.5 and 0.6 ± 0.5 ms for 1 mm, and 0.7 ± 0.5 and 0.5 ± 0.4 ms for 30 mm. C, distributions of duration of the subpause. The distributions were fitted with single-exponential decay functions: constant × exp(−t/τ). The time constants were obtained as 2.6 ± 0.2, 2.7 ± 0.1, 2.1 ± 0.1, 2.4 ± 0.2, 2.3 ± 0.1, and 2.2 ± 0.2 ms for 1 μm, 10 μm, 100 μm, 1 mm, and 30 mm ATP, respectively. The accumulated distribution for all tested [ATP]s is shown at the bottom, and the time constant was estimated as 2.5 ± 0.2 ms. D, distributions of angle difference (Δθ) from main pause to subpause. The distributions were fitted with Gaussian functions. The peak angles were obtained as 36.8 ± 0.4, 32.8 ± 1.8, 41.7 ± 0.4, 42.5 ± 0.3, 42.0 ± 0.6, and 35.2 ± 0.8° at 1 μm, 10 μm, 100 μm, 1 mm, and 30 mm ATP, respectively. The accumulated distribution for all tested [ATP]s is shown at the bottom, and the peak angle was estimated as 39.8 ± 0.1°. E, [ATP] dependences of the time constants. The red circles and blue squares show the time constant of main pause and subpause, respectively. The red line indicates the linear fit of τ1 from 100 nm to 10 μm ATP. F, [ATP] dependence of Δθ. The orange triangles show the peak angle of Gaussian fitting at each [ATP].