Abstract
Multiple cell functions including motility, intracellular transport, and mitosis depend on microtubules. Particularly during mitosis, microtubules must be long and rigid, a quality characterized by persistence length (Lp). Our previous work has shown that the stabilizer GMPCPP increases rigidity (Lp = 1.85 ± 0.5 mm) over Taxol alone (Lp = 0.65 ± 0.1 mm). When under the influence of both GMPCPP and Taxol, the effect of GMPCPP dominates and the microtubule is more rigid (Lp = 1.95 ± 0.7 mm). We investigate microtubules with variable stiffness along their length made from GMPCPP and Taxol segments. Segmented microtubules are polymerized by growing Taxol segments from a GMPCPP seed or annealing segments after each are polymerized separately. Annealing creates defects in the lattice structure, while growth from a seed reduces defects. Two experimental techniques are used to determine persistence length of: (1) individual, freely fluctuating microtubules and (2) microtubules with one fixed end. The use of two methods allows for a more accurate interpretation of the effects of segmentation on microtubule rigidity. We report the initial findings of this study.
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