Kinetic and thermodynamic analyses of outer doublet tubulin polymerization

The kinetics and thermodynamics of microtubule assembly were studied using outer doublet tubulin isolated from starfish sperm. Purified tubulin was gel-filtered just prior to the experiments. The self-polymerization process was found to be characterized by a lag followed by a growth phase that did not show a single first-order decay reaction. This suggested that the nucleation and elongation steps are not completely separate. The maximal rate of polymerization and the reciprocal of the duration of lag time increased approximately in proportion to the third and second powers of tubulin concentration above the critical concentration, respectively. Measurement of temperature dependence of the critical concentration gave a linear van't Hoff plot with delta H degree = 13.5 kcal/mol and delta S degree = 69 e.u.