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. 2018 Jul 10;293(35):13389–13400. doi: 10.1074/jbc.RA118.002767

Figure 3.

Figure 3.

KIF3AB fluorescence transients and simulations. KIF3AB was rapidly mixed in the stopped-flow instrument with varying concentrations of microtubules plus 50 μm mantATP (25 μm mantATP final) to determine the time course of microtubule association and ADP release. A, low microtubule concentration family (1.75–4 μm) at 0.5 μm KIF3AB dimer. B, simulations of the low microtubule concentration family. C, high microtubule concentration family (5–20 μm) at 1 μm KIF3AB dimer. D, simulations of the high microtubule concentration family. E, a double exponential function was also fit to each transient, and the observed rates of the fast component were plotted as a function of microtubule concentration. The hyperbolic fit to the data provided the maximum rate constant of ADP release at 39.6 ± 0.9 s−1 with a K1/2,MT at 3.8 ± 0.3 μm (mean ± S.E.). In simulation panels B and D, simulations are indicated by solid black lines, and only 100 of 1000 data points for each experimental transient are presented for clarity. The green transient in A and B is the buffer control in the absence of microtubules at each KIF3AB concentration.