A novel microtubule-based motor protein (KIF4) for organelle transports, whose expression is regulated developmentally.
A novel microtubule-based motor protein (KIF4) for organelle transports, whose expression is regulated developmentally.
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一种用于细胞器转运的新型微管运动蛋白（KIF4），其表达受到发育的调节。
DOI：
10.1083/jcb.127.1.187
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发表时间：
1994-10
期刊：
影响因子：
--
通讯作者：
Hirokawa N
中科院分区：
文献类型：
--
作者：
Sekine Y;Okada Y;Noda Y;Kondo S;Aizawa H;Takemura R;Hirokawa N
为了了解轴突中细胞器的运输机制，我们从小鼠脑中分离并测序了编码KIF 4的cDNA，并对其进行了生物化学和免疫细胞化学表征。KIF 4的完整氨基酸序列分析和在Sf 9细胞中表达的KIF 4分子的超微结构研究显示，该蛋白质含有1，231个氨基酸残基（M（r）139，550），并且该分子（具有球状头部和尾部的116-nm杆）由三个结构域组成：NH 2-末端球状马达结构域、中央α-螺旋柄结构域和COOH-末端尾部结构域。KIF 4蛋白具有与微管的核苷酸依赖性结合、微管激活的ATP酶活性和微管加末端定向运动的性质。北方印迹分析和原位杂交结果表明，KIF 4在幼年小鼠的组织中有较强的表达，包括分化的幼年神经元，而在成年小鼠中，除脾脏外，其表达显著降低。免疫细胞化学研究表明，KIF 4与膜细胞器共定位在分化的神经元的生长锥和培养的成纤维细胞的细胞质中。在细胞周期的有丝分裂期，KIF 4似乎与有丝分裂纺锤体中的膜性细胞器共定位。因此，我们得出结论，KIF 4是一种新的微管相关的膜细胞器的顺行马达蛋白，其表达的调节发育。
To understand the mechanisms of transport for organelles in the axon, we isolated and sequenced the cDNA encoding KIF4 from murine brain, and characterized the molecule biochemically and immunocytochemically. Complete amino acid sequence analysis of KIF4 and ultrastructural studies of KIF4 molecules expressed in Sf9 cells revealed that the protein contains 1,231 amino acid residues (M(r) 139,550) and that the molecule (116-nm rod with globular heads and tail) consists of three domains: an NH2-terminal globular motor domain, a central alpha-helical stalk domain and a COOH-terminal tail domain. KIF4 protein has the property of nucleotide-dependent binding to microtubules, microtubule- activated ATPase activity, and microtubule plus-end-directed motility. Northern blot analysis and in situ hybridization demonstrated that KIF4 is strongly expressed in juvenile tissues including differentiated young neurons, while its expression is decreased considerably in adult mice except in spleen. Immunocytochemical studies revealed that KIF4 colocalized with membranous organelles both in growth cones of differentiated neurons and in the cytoplasm of cultured fibroblasts. During mitotic phase of cell cycle, KIF4 appears to colocalize with membranous organelles in the mitotic spindle. Hence we conclude that KIF4 is a novel microtubule-associated anterograde motor protein for membranous organelles, the expression of which is regulated developmentally.