INVITRO ASSEMBLY OF FLAGELLAR OUTER DOUBLET TUBULIN
INVITRO ASSEMBLY OF FLAGELLAR OUTER DOUBLET TUBULIN
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DOI：
10.1083/jcb.79.2.500
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发表时间：
1978-01-01
影响因子：
7.8
通讯作者：
ROSENBAUM, JL
中科院分区：
文献类型：
--
作者：
BINDER, LI;ROSENBAUM, JL
鞭毛外双微管[来自海胆Strongylocentrotus drobachiensis或S. purpuratus]的微管蛋白在体外重组为含有14和15个原丝的单个微管。微管蛋白组装依赖于KCl和微管蛋白浓度，在最佳溶剂条件下表现出0.72 mg/ml的临界浓度。鞭毛微管蛋白的纯化循环的温度依赖性组装-拆卸和分子筛层析，其特征在于通过二维凝胶电泳。虽然在体外没有形成双微管，外部双微管蛋白组装到完整的A-和B-亚纤维的外部双微管和基体的赖氏衣原体;从这些结构的远端组装的速率大于从近端。来自小牛脑的微管相关蛋白刺激外部双重微管蛋白组装，用细丝状突起装饰微管。
Flagellar outer doublet microtubules [from sperm of the sea urchin Strongylocentrotus drobachiensis or S. purpuratus] were solubilized by use of sonication and the tubulin was reassembled in vitro into single microtubules conaining 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 2-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 reinhardii; the rate of assembly from the distal ends of these structures was greater than that from the proximal ends. Microtubule-associated proteins from calf brain stimulated outer doublet tubulin assembly, decorating the microtubules with fine filamentous projections.