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首页> 外文期刊>Molecular and Cellular Biology >Wiskott-Aldrich syndrome protein physically associates with Nck through Src homology 3 domains.
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Wiskott-Aldrich syndrome protein physically associates with Nck through Src homology 3 domains.

机译：Wiskott-Aldrich综合征蛋白通过Src同源性3结构域与Nck物理结合。

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In the second of a series of experiments designed to identify p47nck-Src homology 3 (SH3)-binding molecules, we report the cloning of SAKAP II (Src A box Nck-associated protein II) from an HL60 cDNA expression library. This molecule has been identified as a cDNA encoding the protein product of WASP, which is mutated in Wiskott-Aldrich syndrome patients. Studies in vivo and in vitro demonstrated a highly specific interaction between the SH3 domains of p47nck and Wiskott-Aldrich syndrome protein. Furthermore, anti-Wiskott-Aldrich syndrome protein antibodies recognized a protein of 66 kDa by Western blot (immunoblot) analysis. In vitro translation studies identified the 66-kDa protein as the protein product of WASP, and subcellular fractionation experiments showed that p66WASP is mainly present in the cytosol fraction, although significant amounts are also present in membrane and nuclear fractions. The main p47nck region implicated in the association with p66WASP was found to be the carboxy-terminal SH3 domain.
机译：在旨在鉴定p47nck-Src同源性3（SH3）结合分子的一系列实验的第二步中，我们报道了从HL60 cDNA表达文库克隆SAKAP II（与Src A盒Nck相关的蛋白II）。该分子已被鉴定为编码WASP蛋白产物的cDNA，该蛋白在Wiskott-Aldrich综合征患者中发生了突变。体内和体外研究表明，p47nck的SH3结构域与Wiskott-Aldrich综合征蛋白之间具有高度特异性的相互作用。此外，通过Western blot（免疫印迹）分析，抗维斯克特-奥尔德里奇综合症蛋白抗体识别出66 kDa的蛋白。体外翻译研究将66 kDa蛋白鉴定为WASP的蛋白质产物，亚细胞分级分离实验表明p66WASP主要存在于胞质溶胶级分中，尽管在膜和核级分中也大量存在。发现与p66WASP有关的主要p47nck区域是羧基末端SH3结构域。

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