Cell Structure and Function
Online ISSN : 1347-3700
Print ISSN : 0386-7196
ISSN-L : 0386-7196
Removal of βIII Isotype Enhances Taxol Induced Microtubule Assembly
Qing LuRichard F. Luduena
Author information
JOURNALS FREE ACCESS

1993 Volume 18 Issue 3 Pages 173-182

Details
Abstract

The interaction of βIII-depleted tubulin with taxol was investigated. A monoclonal antibody against the βIII tubulin isotype was immobilized on a sepharose 4B column and used to remove the βIII tubulin isotype from unfractionated tubulin. The assembly of βIII-depleted tubulin in the presence of taxol was enhanced compared to unfractionated tubulin. The critical concentration of unfractionated tubulin in the presence of 10 μM taxol is 0.4 mg/ml, while the critical concentration of βIII-depleted tubulin is 0.16 mg/ml. At different concentrations of taxol, the assembly of βIII-depleted tubulin is increased relative to that of un fractionated tubulin and reaches the maximumat about a 1 : 1 ratio of tubulin and taxol. The assembly of un fractionated tubulin and βIII-depleted tubulin has also been studied by electron microscopy. After 2 minutes at 37°C, unfractionated tubulin assembly in the presence of 10 μM taxol results only in ribbon-like and ring structures; there are no visible microtubules. By 5 minutes, microtubules appear and increase in length. The assembly of βIII-depleted tubulin in the presence of 10 μM taxol occurs more quickly. In contrast to the case with un fractionated tubulin, βIII-depleted tubulin assembles within 2 minutes into microtubules which increase in length with time. At 30 minutes, microtubules assembled from βIII-depleted tubulin are shorter than the microtubules assembled from unfractionated tubulin. There is no visible difference between the microtubules assembled from unfractionated tubulin and βIII-depleted tubulin. Taxol-induced βIII-depleted tubulin assembly is more resistant to the inhibiting effect of podophyllotoxin and colchicine. It is also less sensitive to the inhibiting effect of cold temperature.

Information related to the author
© Japan Society for Cell Biology
Previous article Next article
feedback
Top