Removal of βIII Isotype Enhances Taxol Induced Microtubule Assembly

Cell Structure and Function

Richard F. Luduena¹  Qing Lu¹ 

[1] Department of biochemistry, The University of Texas Health Science Center at San Antonio

关键词: β-tubulin isotypes;    microtubule assembly;    taxol;   

DOI  :  10.1247/csf.18.173

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(30)Cited-By(39)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.

【 授权许可】

Unknown