鞭毛外双联微管蛋白的体外组装
The in vitro assembly of flagellar outer doublet tubulin.
作者信息
Binder L I, Rosenbaum J L
出版信息
J Cell Biol. 1978 Nov;79(2 Pt 1):500-15. doi: 10.1083/jcb.79.2.500.
Abstract
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.
摘要
鞭毛外双联体微管通过超声处理使其溶解，微管蛋白在体外重新组装成含有14和15条原纤维的单微管。微管蛋白组装依赖于KCl和微管蛋白浓度，在最佳溶剂条件下临界浓度为0.72mg/ml。通过温度依赖性组装-拆卸循环和分子筛色谱法纯化鞭毛微管蛋白，并通过二维凝胶电泳进行表征。尽管体外未形成双联体微管，但外双联体微管蛋白组装到衣藻外双联体微管完整的A亚纤维和B亚纤维以及基体上；这些结构远端的组装速率大于近端。来自哺乳动物脑的微管相关蛋白（MAPs）刺激外双联体微管蛋白组装，用细丝状突起装饰微管。
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