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爪蟾卵母细胞中一种微管组装抑制剂的特性分析

Characterization of a microtubule assembly inhibitor from Xenopus oocytes.

作者信息

Govindan B, Vale R D

机构信息

Department of Cellular and Molecular Pharmacology, University of California, San Francisco 94143-0450, USA.

出版信息

Cell Motil Cytoskeleton. 2000 Jan;45(1):51-7. doi: 10.1002/(SICI)1097-0169(200001)45:1<51::AID-CM5>3.0.CO;2-7.

Abstract

The dynamic properties of microtubules (MTs) are important for a wide variety of cellular processes, including cell division and morphogenesis. MT assembly and disassembly in vivo are regulated by cellular factors that influence specific parameters of MT dynamics. Here, we describe the characterization of a previously reported MT assembly inhibitor activity from Xenopus oocytes [Gard and Kirschner, 1987: J. Cell Biol. 105:2191-2201]. Video microscopy measurements reveal that the inhibitor specifically decreases the plus end growth rate of MTs and increases the critical concentration for tubulin. However, catastrophe frequency, rescue frequency, and shrinkage rates are not affected by the activity. Chromatography on Mono Q and hydroxyapatite columns has shown that the activity cofractionates with a subpopulation of tubulin. This tubulin subpopulation and the MT assembly inhibitor activity also co-migrate with a large S value (25-30S) on sucrose gradients. The high molecular weight tubulin complex and the MT assembly inhibitor activity are both developmentally regulated and disappear after oocyte maturation with progesterone.

摘要

微管(MTs)的动态特性对于包括细胞分裂和形态发生在内的多种细胞过程都很重要。体内微管的组装和拆卸受影响微管动力学特定参数的细胞因子调控。在此,我们描述了非洲爪蟾卵母细胞中先前报道的一种微管组装抑制剂活性的特征[加德和基尔希纳,1987年:《细胞生物学杂志》105:2191 - 2201]。视频显微镜测量显示,该抑制剂特异性降低微管的正端生长速率,并提高微管蛋白的临界浓度。然而,灾难频率、拯救频率和收缩速率不受该活性影响。在单Q柱和羟基磷灰石柱上进行的色谱分析表明,该活性与微管蛋白的一个亚群共分离。这个微管蛋白亚群和微管组装抑制剂活性在蔗糖梯度上也与一个大的S值(25 - 30S)一起迁移。高分子量微管蛋白复合物和微管组装抑制剂活性在发育过程中都受到调控,并且在卵母细胞用孕酮成熟后消失。

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