LIM kinase 1, a key regulator of actin dynamics, is widely expressed in embryonic and adult tissues
LIM kinase 1, a key regulator of actin dynamics, is widely expressed in embryonic and adult tissues
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DOI：
10.1016/j.yexcr.2003.11.024
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发表时间：
2004-04-01
影响因子：
3.7
通讯作者：
Bernard, O
中科院分区：
文献类型：
--
作者：
Foletta, VC;Moussi, N;Bernard, O
内源性LIM激酶1（LIMK1）蛋白的表达进行了研究，在胚胎和成年小鼠使用大鼠单克隆抗体（mAb），它识别特异性的PDZ结构域的LIMK1，而不是LIMK2。免疫印迹分析显示LIMK1在组织和细胞系中以70-kDa蛋白的形式广泛表达，在一些组织和细胞系中存在更高质量的形式（约75 kDa）。约50 kDa的较小亚型偶尔也很明显。免疫荧光分析表明，LIMK1的亚细胞定位在成纤维细胞中的粘着斑，除了核周（高尔基体）和偶尔的核定位与肌动蛋白，桩蛋白和黏着斑蛋白共染色所揭示的。此外，LIMK1和桩蛋白之间的关联，而不是黏着斑蛋白被确定通过免疫共沉淀分析。LIMK1在E18大鼠海马锥体神经元的轴突和树突生长锥中富集，其中在延伸到丝状伪足的点中以及在被鉴定为高尔基体的核周区域中发现LIMK1。在原位，我们确定LIMK1蛋白表达在所有胚胎和成人组织检查，虽然在不同的水平和不同的细胞群。大鼠单克隆LIMK1抗体识别来自许多物种的细胞和组织提取物中类似大小的蛋白质。因此，LIMK I是一种广泛表达的蛋白质，以几种同种型存在。(C)2004年爱思唯尔公司All rights reserved.
The expression of endogenous LIM kinase 1 (LIMK1) protein was investigated in embryonic and adult mice using a rat monoclonal antibody (mAb), which recognizes specifically the PDZ domain of LIMK1 and not LIMK2. Immunoblotting analysis revealed widespread expression of LIMK1 existing as a 70-kDa protein in tissues and in cell lines, with a higher mass form (approximately 75 kDa) present in some tissues and cell lines. Smaller isoforms of approximately 50 kDa were also occasionally evident. Immunofluorescence analysis demonstrated LIMK1 subcellular localization at focal adhesions in fibroblasts as revealed by co-staining with actin, paxillin and vinculin in addition to perinuclear (Golgi) and occasional nuclear localization. Furthermore, an association between LIMK1 and paxillin but not vinculin was identified by co-immunoprecipitation analysis. LIMK1 is enriched in both axonal and dendritic growth cones of E18 rat hippocampal pyramidal neurons where it is found in punctae that extend far out into filopodia, as well as in a perinuclear region identified as Golgi. In situ, we identify LIMK1 protein expression in all embryonic and adult tissues examined, albeit at different levels and in different cell populations. The rat monoclonal LIMK1 antibody recognizes proteins of similar size in cell and tissue extracts from numerous species. Thus, LIMK I is a widely expressed protein that exists as several isofonns. (C) 2004 Elsevier Inc. All rights reserved.