Mechanism of tubulin assembly: guanosine 5'-triphosphate hydrolysis decreases the rate of microtubule depolymerization.

The rate of depolymerization of microtubules upon lowering the temperature was found to depend on the amount of time elapsed since the beginning of the assembly process. In the first minutes following self-assembly at 37 degrees C, microtubules are more cold sensitive and depolymerize faster than later at the steady state. In the meanwhile, no change occurred in the average length nor in the shape of the distribution of microtubules. On the other hand, the evolution with time of the apparent dissociation rate constant of tubulin from microtubules was in good correlation with the GTP content of microtubules following assembly, showing that GTP hydrolysis modifies the tubulin-tubulin interactions. Microtubule-bound GTP was not exchangeable for GDP, but steady-state GTP hydrolysis was inhibited by GDP. This result indicates that GDP and GTP exhibit different affinities for tubulin in the body and at the ends of microtubules. It is proposed that GTP-tubulin dissociates faster from microtubules than GDP-tubulin. In other words GTP hydrolysis contributes to the stabilization of microtubules.