Figure 6. MT•Kar3Vik1 interactions upon neck coiled coil cross-linking. (A) Model of Kar3Vik1 identifying the sites on Kar3 E382C and Vik1 K372C of the 10-Å and 20-Å cross-links. See Fig. S5 for cross-linking efficiency. (B) Chemical structures of M4M (10-Å cross-link) and DPDPB (20-Å cross-link). (C) Histograms of the velocity distribution of gliding microtubules for WT GCN4-Kar3Vik1, non-cross-linked, 10-Å cross-linked, and 20-Å cross-linked Kar3Vik1 motors. The rates are presented in 0.25 µm/min bins with the mean speed reported as mean ± SEM; n = number of microtubules scored. The data shown represent a total of 12–25 movies for each specific motor recorded on 3–6 different days. Representative movies are included as supplemental material. (D) Analysis of the persistence of motility promoted by non-cross-linked, 20-Å cross-linked, and 10-Å cross-linked GCN4-Kar3Vik1. Gliding microtubules were tracked for the initial 20 min of the experiments, and the percentage of microtubules that continued to move is presented: n = 110 microtubules for non-cross-linked, 110 microtubules for 20-Å cross-linked, and 221 microtubules for 10-Å cross-linked GCN4-Kar3Vik1. (E) MT•Kar3Vik1 cosedimentation to determine stoichiometry of binding. Gel images are not shown.
Figure 6.

MT•Kar3Vik1 interactions upon neck coiled coil cross-linking. (A) Model of Kar3Vik1 identifying the sites on Kar3 E382C and Vik1 K372C of the 10-Å and 20-Å cross-links. See Fig. S5 for cross-linking efficiency. (B) Chemical structures of M4M (10-Å cross-link) and DPDPB (20-Å cross-link). (C) Histograms of the velocity distribution of gliding microtubules for WT GCN4-Kar3Vik1, non-cross-linked, 10-Å cross-linked, and 20-Å cross-linked Kar3Vik1 motors. The rates are presented in 0.25 µm/min bins with the mean speed reported as mean ± SEM; n = number of microtubules scored. The data shown represent a total of 12–25 movies for each specific motor recorded on 3–6 different days. Representative movies are included as supplemental material. (D) Analysis of the persistence of motility promoted by non-cross-linked, 20-Å cross-linked, and 10-Å cross-linked GCN4-Kar3Vik1. Gliding microtubules were tracked for the initial 20 min of the experiments, and the percentage of microtubules that continued to move is presented: n = 110 microtubules for non-cross-linked, 110 microtubules for 20-Å cross-linked, and 221 microtubules for 10-Å cross-linked GCN4-Kar3Vik1. (E) MT•Kar3Vik1 cosedimentation to determine stoichiometry of binding. Gel images are not shown.

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