IDENTIFICATION AND LOCALIZATION OF HUNTINGTIN IN BRAIN AND HUMAN LYMPHOBLASTOID CELL-LINES WITH ANTI-FUSION PROTEIN ANTIBODIES
IDENTIFICATION AND LOCALIZATION OF HUNTINGTIN IN BRAIN AND HUMAN LYMPHOBLASTOID CELL-LINES WITH ANTI-FUSION PROTEIN ANTIBODIES
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DOI：
10.1073/pnas.92.19.8710
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发表时间：
1995-09-12
影响因子：
11.1
通讯作者：
HERSCH, SM
中科院分区：
文献类型：
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作者：
GUTEKUNST, CA;LEVEY, AI;HERSCH, SM
亨廷顿病 (HD) 表型与 IT15 基因中三核苷酸重复的扩展有关，该基因预计编码一种名为亨廷顿蛋白的 348 kDa 蛋白质。我们使用多克隆和单克隆抗融合蛋白抗体来鉴定大鼠、猴子和人类中的天然亨廷顿蛋白。蛋白质印迹显示具有预期分子量的蛋白质存在于大鼠和猴脑组织以及对照病例的淋巴母细胞系的可溶性部分中。在青少年发病的杂合子 HD 病例的淋巴母细胞系中，正常和突变亨廷顿蛋白均表达，并且增加重复扩增导致突变蛋白水平降低。免疫化学表明亨廷顿蛋白存在于整个大脑的神经元中，在较大的神经元中水平最高。在人类纹状体中，亨廷顿蛋白以斑块状分布富集，可能对应于 HD 受影响的第一个区域。亨廷顿蛋白的亚细胞定位与主要在体细胞树突区域中发现的胞质蛋白一致。亨廷顿蛋白似乎特别与微管相关，尽管有些也与突触小泡相关。根据亨廷顿蛋白与微管相关的定位，我们推测该突变损害了线粒体、囊泡或其他细胞器或分子的细胞骨架锚定或运输。
The Huntington disease (HD) phenotype is associated with expansion of a trinucleotide repeat in the IT15 gene, which is predicted to encode a 348-kDa protein named huntingtin. We used polyclonal and monoclonal anti-fusion protein antibodies to identify native huntingtin in rat, monkey, and human. Western blots revealed a protein with the expected molecular weight which is present in the soluble fraction of rat and monkey brain tissues and lymphoblastoid cell lines from control cases. In lymphoblastoid cell lines from juvenile-onset heterozygote HD cases, both normal and mutant huntingtin are expressed, and increasing repeat expansion leads to lower levels of the mutant protein. Immunocgtochemistry indicates that huntingtin is located in neurons throughout the brain, with the highest levels evident in larger neurons. In the human striatum, huntingtin is enriched in a patch-like distribution, potentially corresponding to the first areas affected in HD. Subcellular localization of huntingtin is consistent with a cytosolic protein primarily found in somatodendritic regions. Huntingtin appears to particularly associate with microtubules, although some is also associated with synaptic vesicles. On the basis of the localization of huntingtin in association with microtubules, we speculate that the mutation impairs the cytoskeletal anchoring or transport of mitochondria, vesicles, or other organelles or molecules.