Linkages between the effects of taxol, colchicine, and GTP on tubulin polymerization.
A comparative study has been carried out of the effects of taxol on the polymerizations into microtubules of microtubule-associated protein-free tubulin, prepared by the modified Weisenberg procedure, and of the tubulin-colchicine complex into large aggregates. Taxol enhances, to a much greater extent, the stability of microtubules than that of the tubulin-colchicine polymers so that, with highly purified tubulin, assembly into microtubules takes place at 10 degrees C, even in the absence of exogenous GTP. The polymerization of tubulin-colchicine requires both heat and GTP, and the process is reversed by cooling. These results indicate that in both systems polymerization is linked to interactions with taxol and GTP, the interplay of linkage free energies imparting the observed polymer stabilities. In the case of microtubule formation, the linkage free energy provided by taxol binding is approximately -3.0 kcal/mol of alpha-beta-tubulin dimer, whereas this quantity is reduced to approximately -0.5 kcal/mol in tubulin-colchicine, indicating the expenditure of much more binding free energy in the latter case for overcoming unfavorable factors, such as steric hindrance and geometric strain. The difference in the effect of GTP on the two polymerization processes reflects the respective abilities of the bindings of taxol to the two states of tubulin to overcome the loss of the linkage free energy of GTP binding. Analysis of the linkages leads to the conclusions that taxol need not change qualitatively the mechanism of microtubule assembly and that tubulin with the E-site unoccupied by nucleotide should have the capacity to form microtubules, the reaction being extremely weak.
D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D003078:Colchicine / Q000494:pharmacology*; D006160:Guanosine Triphosphate / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000494:pharmacology; D008870:Microtubules / Q000187:drug effects; D017239:Paclitaxel; D011108:Polymers / Q000378:metabolism; D013816:Thermodynamics; D014404:Tubulin / Q000378:metabolism*
D000470:Alkaloids; D011108:Polymers; D014404:Tubulin; D006160:Guanosine Triphosphate; D008274:Magnesium; D017239:Paclitaxel; D003078:Colchicine