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首页> 外文期刊>Molecular and Cellular Biology >Protein 4.1 R-135 Interacts with a Novel Centrosomal Protein (CPAP) Which Is Associated with the γ-Tubulin Complex
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Protein 4.1 R-135 Interacts with a Novel Centrosomal Protein (CPAP) Which Is Associated with the γ-Tubulin Complex

机译：蛋白质4.1 R-135与与γ-微管蛋白复合物相关的新型中心体蛋白（CPAP）相互作用

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Using a yeast two-hybrid system, we isolated a novel human centrosomal protein, CPAP (centrosomal P4.1-associated protein), which specifically interacts with the head domain of the 135-kDa protein 4.1R isoform (4.1R-135). Sequence analysis revealed that the carboxyl terminus of CPAP has 31.3% amino acid identity with human Tcp-10 (at-complex responder gene product). Interestingly, most of the sequence identity is restricted to two conserved regions. One carries a leucine zipper, which may form a series of heptad repeats involved in coiled-coil formation; the other contains unusual glycine repeats with unknown function. Immunofluorescence analysis revealed that CPAP and γ-tubulin are localized within the centrosome throughout the cell cycle. CPAP cosediments with γ-tubulin in sucrose gradients and coimmunoprecipitates with γ-tubulin, indicating that CPAP is a part of the γ-tubulin complex. Furthermore, functional analysis revealed that CPAP is localized within the center of microtubule asters and may participate in microtubule nucleation. The formation of microtubule asters was significantly inhibited by anti-CPAP antibody. Together, these observations indicate that CPAP may play an important role in cell division and centrosome function.
机译：使用酵母双杂交系统，我们分离了一种新型的人类中心体蛋白CPAP（与中心体P4.1相关的蛋白），该蛋白与135-kDa蛋白4.1R亚型（4.1R-135）的头部结构域特异性相互作用。序列分析表明，CPAP的羧基末端与人Tcp-10（a t -复杂应答基因产物）具有31.3％的氨基酸同一性。有趣的是，大多数序列同一性仅限于两个保守区。一个带有亮氨酸拉链，它可能形成一系列参与卷曲螺旋形成的七肽重复序列;另一个包含功能未知的异常甘氨酸重复序列。免疫荧光分析表明，CPAP和γ-微管蛋白在整个细胞周期内都位于中心体中。 CPAP在蔗糖梯度中与γ-微管蛋白共沉淀，并与γ-微管蛋白共免疫沉淀，表明CPAP是γ-微管蛋白复合物的一部分。此外，功能分析表明，CPAP位于微管紫苑的中心，并且可能参与微管成核。抗CPAP抗体显着抑制了微管紫苑的形成。这些观察结果共同表明，CPAP可能在细胞分裂和中心体功能中发挥重要作用。

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