MAP1B and Clathrin Are Novel Interacting Partners of the Giant Cyto-linker Dystonin
MAP1B and Clathrin Are Novel Interacting Partners of the Giant Cyto-linker Dystonin
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DOI：
10.1021/pr200564g
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发表时间：
2011-11-01
影响因子：
4.4
通讯作者：
Kothary, Rashmi
中科院分区：
文献类型：
--
作者：
Bhanot, Kunal;Young, Kevin G.;Kothary, Rashmi
肌张力障碍素是一种在神经系统中起重要作用的大的多结构域细胞内相关蛋白。肌张力障碍蛋白的缺失导致神经肌肉功能障碍和小鼠突变体肌张力障碍的早期死亡。在相关蛋白质中保守，斑蛋白结构域是所有主要肌张力障碍蛋白亚型的定义特征，但其相互作用尚未详细探讨。本研究的目的是确定新的相互作用的合作伙伴的斑域的神经亚型肌张力障碍（肌张力障碍-a）。新发现的相互作用的蛋白质通过下拉试验进行了验证，使用coimmunoprecipitation，coimmunofluorescence，和邻近连接试验。微管相关蛋白1B（MAP 1B），一种微管稳定蛋白，和网格蛋白重链，网格蛋白三聚体的主要成分，被确定为肌张力蛋白-a的相互作用伴侣。磷酸化MAP 1B水平的增加表明MAP 1B的失调和dt病理学的潜在新组分。这项工作将进一步促进我们对细胞骨架蛋白如何影响和调节神经退行性疾病的理解。
Dystonin is a large multidomain cytoskeletal-associated protein that plays an essential role in the nervous system. Loss of dystonin results in neuromuscular dysfunction and early death in a mouse mutant called dystonia musculorum. Conserved among related proteins, the plakin domain is a defining feature of all major dystonin isoforms, yet its interactions have not been explored in detail. The purpose of the present study was to identify novel interacting partners of the plakin domain of the neuronal isoform of dystonin (dystonin-a). Newly identified interacting proteins discovered through a pull-down assay were validated using coimmunoprecipitation, coimmunofluorescence, and proximity ligation assays. Microtubule associated protein 1B (MAP1B), a microtubule stabilizing protein, and clathrin heavy chain, the major component of the clathrin triskelion, were identified as interaction partners for dystonin-a. Increased levels of phosphorylated MAP1B suggest a misregulation of MAP1B and a potentially novel component of the dt pathology. This work will further facilitate our understanding of how cytoskeletal proteins can affect and regulate neurodegenerative disorders.