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[Preprint]. 2023 Sep 20:2023.01.26.525761. Originally published 2023 Jan 27. [Version 3] doi: 10.1101/2023.01.26.525761

Figure 5. Disruption of the hierarchical folding activates KIF5B motility in vitro.

Figure 5.

(A) The designed KIF5B mutations and truncations shown on the domain diagram. (B) Example kymographs of KIF5B with different truncations and the corresponding landing rate. Landing rate (Events/μm/s/μM): Lines show mean ± SEM: 1.07 ± 0.16 (full length), 14.92 ± 1.30 (1–909), 15.75 ± 0.70 (1–890), 26.87 ± 1.65 (1–565), 53.42 ± 3.47 (1–420). N=2; n=25, 20, 23, 15 and 14 MTs. One-way ANOVA followed by Dunnett’s test. ****P<0.0001. (C) Example kymographs of KIF5B with single mutation and the corresponding landing rate. Landing rate (Events/μm/s/μM): Lines show mean ± SEM: 1.07 ± 0.16 (full length), 3.93 ± 0.32 (CC2), 4.93 ± 0.42 (Hinge), 6.90 ± 0.78 (IAK), 0.67 ± 0.32 (Linker). N=2; n=25, 25, 25, 20, and 9 MTs. One-way ANOVA followed by Dunnett’s test. *P<0.05, ***P<0.001, n.s not significant. (D) Example kymographs of KIF5B with combined mutations and the corresponding landing rate. Landing rate (Events/μm/s/μM): Lines show mean ± SEM: 1.07 ± 0.16 (full length), 6.44 ± 0.40 (CC2+hinge), 7.66 ± 0.86 (CC2+IAK), 9.65 ± 0.85 (Hinge+IAK), 10.49 ± 0.57 (CC2+hinge+IAK). N=2; n=25, 15, 12, 20, and 32 MTs. One-way ANOVA followed by Dunnett’s test. **P<0.01, ***P<0.001, ****P<0.0001.