In Vitro Reconstitution of Calf Brain Microtubules: Effects of Macromolecules

A comparative study has been carried out of the ability to reconstitute into microtubules of tubulins prepared by the Weisenberg and the cycle procedures. It was found that further purification of cycle tubulin by phosphocellulose chromatography made its ability to polymerize identical with that of Weisenberg tubulin. By adding to either tubulin the isolated proteins which copurify with tubulin in the cycle procedure, it is possible to reduce their critical concentrations of microtubule formation to a value identical with that of cycletubulin. It was demonstrated quantitatively that the effect of these nontubulin proteins could be mimicked by a variety of polycationic molecules, the most effective one being poly(Llysine). A possible mechanism is described by which growing microtubules could be stabilized by subsequent addition of the nontubulin proteins. The conclusion is drawn that, at present, it is not known whether the proteins which copurify with tubulin are specific components of the microtubule system, or simply artefactually coprecipitated impurities.