Substructural analysis of the microtubule and its polymorphic forms.

Microtubules are composed of 13 protofilaments which are aligned parallel to the long axis of the tubule, as can be seen in thin sections of material fixed in the presence of tannic acid. Evidence is presented based upon Markham rotations of the isolated flagellar axoneme that the bridges (spoke, nexin, dynein) must connect to certain specified protofilaments. By means of geometric considerations it is concluded that the bridges in a Centrohelida and Raphidiophrys also connect certain specified protofilaments. Two polymorphic forms of microtubules were also examined using tannic acid in the fixative, a 320 A tubule produced in Echinosphaerium by treatment with low temperature, and a vinblastine-induced microtubule crystal. In both forms, the arrangement of the subunits is not in the form of linear protofilaments. The vinblastine crystal in transverse section is made up of hexagonally packed tubules approximately 320 A in diameter. The wall of each tubule in turn is composed of two parallel protofilaments which spiral upwards, giving the appearance of a barber pole. The center-to-center separation of adjacent protofilaments is 180 A. We conclude that the tubulin heterodimer lies parallel to the axis of the protofilaments, exposing the colchicine binding site.