The in vitro assembly of flagellar outer doublet tubulin
The in vitro assembly of flagellar outer doublet tubulin
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鞭毛外双联微管蛋白的体外组装
DOI：
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发表时间：
1978
影响因子：
7.8
通讯作者：
J. Rosenbaum
中科院分区：
文献类型：
--
作者：
L. Binder;J. Rosenbaum
通过超声处理溶解鞭毛外部双联微管，微管蛋白在体外重新组装成含有 14 和 15 个原丝的单个微管。微管蛋白组装取决于 KCl 和微管蛋白浓度，在最佳溶剂条件下表现出 0.72 mg/ml 的临界浓度。通过温度依赖性组装-拆卸和分子筛色谱循环纯化鞭毛微管蛋白，并通过二维凝胶电泳进行表征。尽管双联微管未在体外形成，但外双联微管蛋白组装到外双联微管和衣藻基体的完整A-和B-亚纤维上；从这些结构的远端组装的速率大于从近端组装的速率。来自哺乳动物大脑的微管相关蛋白（MAP）刺激外部双联微管蛋白组装，用细丝状突起装饰微管。
Flagellar outer doublet microtubules were solubilized by use of sonication, and the tubulin was reassembled in vitro into single microtubules containing 14 and 15 protofilaments. The tubulin assembly was dependent on both the KCl and tubulin concentrations, exhibiting a critical concentration of 0.72 mg/ml at optimum solvent conditions. Flagellar tubulin was purified by cycles of temperature-dependent assembly-disassembly and molecular sieve chromatography, and characterized by two-dimensional gel electrophoresis. Although doublet microtubules were not formed in vitro, outer doublet tubulin assembled onto intact A- and B-subfibers of outer doublet microtubules and basal bodies of Chlamydomonas; the rate of assembly from the distal ends of these structures was greater than that from the proximal ends. Microtubule-associated proteins (MAPs) from mammalian brain stimulated outer doublet tubulin assembly, decorating the microtubules with fine filamentous projections.