Characterization of palladin, a novel protein localized to stress fibers and cell adhesions.
Characterization of palladin, a novel protein localized to stress fibers and cell adhesions.
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DOI：
10.1083/jcb.150.3.643
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发表时间：
2000-08-07
期刊：
影响因子：
--
通讯作者：
Otey CA
中科院分区：
文献类型：
--
作者：
Parast MM;Otey CA
在这里，我们描述了一种名为palladin的新型磷酸化蛋白的鉴定，该蛋白与α-肌动蛋白共定位于应力纤维、局灶粘连、细胞-细胞连接和胚胎z线。Palladin在成纤维细胞中以90-92-kD双偶体表达，在成纤维细胞裂解物中以α-肌动蛋白复合物的形式共免疫沉淀。利用单克隆抗体筛选小鼠胚胎文库，分离到一个编码palladin的cDNA。Palladin在分子的nh2末端有一个富含脯氨酸的区域，在cooh末端有三个串联的Ig C2结构域。在鸡和小鼠组织的北部和西部印迹中，检测到多种帕拉丁同工型。Palladin在胚胎组织中普遍表达，在小鼠的某些成年组织中表达下调。采用Rcho-1滋养细胞模型研究钯素在体外培养细胞中的作用。当Rcho-1细胞开始组装应力纤维时，观察到Palladin的表达增加。在Rcho-1细胞和成纤维细胞中，使用反义结构物来减弱palladin的表达，并在两种细胞类型中观察到细胞骨架的破坏。在较长时间的反义处理后，成纤维细胞变得完全圆润。这些结果表明，钯是肌动蛋白细胞骨架和局灶黏附的正常组织所必需的。
Here, we describe the identification of a novel phosphoprotein named palladin, which colocalizes with α-actinin in the stress fibers, focal adhesions, cell–cell junctions, and embryonic Z-lines. Palladin is expressed as a 90–92-kD doublet in fibroblasts and coimmunoprecipitates in a complex with α-actinin in fibroblast lysates. A cDNA encoding palladin was isolated by screening a mouse embryo library with mAbs. Palladin has a proline-rich region in the NH2-terminal half of the molecule and three tandem Ig C2 domains in the COOH-terminal half. In Northern and Western blots of chick and mouse tissues, multiple isoforms of palladin were detected. Palladin expression is ubiquitous in embryonic tissues, and is downregulated in certain adult tissues in the mouse. To probe the function of palladin in cultured cells, the Rcho-1 trophoblast model was used. Palladin expression was observed to increase in Rcho-1 cells when they began to assemble stress fibers. Antisense constructs were used to attenuate expression of palladin in Rcho-1 cells and fibroblasts, and disruption of the cytoskeleton was observed in both cell types. At longer times after antisense treatment, fibroblasts became fully rounded. These results suggest that palladin is required for the normal organization of the actin cytoskeleton and focal adhesions.
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64.5
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