Immunochemical characterization on pathological oligomers of mutant Cu/Zn-superoxide dismutase in amyotrophic lateral sclerosis.
Immunochemical characterization on pathological oligomers of mutant Cu/Zn-superoxide dismutase in amyotrophic lateral sclerosis.
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DOI：
10.1186/s13024-016-0145-9
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发表时间：
2017-01-05
影响因子：
15.1
通讯作者：
Furukawa Y
中科院分区：
文献类型：
--
作者：
Tokuda E;Anzai I;Nomura T;Toichi K;Watanabe M;Ohara S;Watanabe S;Yamanaka K;Morisaki Y;Misawa H;Furukawa Y
Cu/Zn-超氧化物歧化酶（SOD 1）基因的显性突变导致家族性肌萎缩侧索硬化症（SOD 1-ALS），其中错误折叠的SOD 1蛋白作为细胞内包涵体在脊髓运动神经元中积累。通过异常二硫键交联的SOD 1寡聚化已被认为是突变型SOD 1中发生的错误折叠途径之一;然而，在SOD 1-ALS病例中这种寡聚化的病理相关性仍然不清楚。我们在体外制备了专门识别通过二硫键交联的SOD 1寡聚体的抗体。通过使用这些抗体，主要对表达突变SOD 1蛋白的转基因小鼠和人SOD 1-ALS病例的组织样品进行免疫组织化学检查和ELISA。我们显示了我们的抗体的识别特异性，专门对二硫键交联的SOD 1寡聚体通过ELISA使用各种形式的纯化的SOD 1蛋白在体外构象不同的状态。此外，这些抗体的表位被掩埋在SOD 1的天然折叠结构中，并且不可接近。然后发现抗体特异性检测SOD 1-ALS患者以及转基因模型小鼠的脊髓运动神经元中的病理性SOD 1种类。我们的研究结果表明，通过二硫键交联的SOD 1寡聚化与SOD 1结构内部的暴露有关，是SOD 1-ALS病例中发生的病理过程。本文的在线版本（doi：10.1186/s13024-016-0145-9）包含补充材料，可供授权用户使用。
Dominant mutations in Cu/Zn-superoxide dismutase (SOD1) gene cause a familial form of amyotrophic lateral sclerosis (SOD1-ALS) with accumulation of misfolded SOD1 proteins as intracellular inclusions in spinal motor neurons. Oligomerization of SOD1 via abnormal disulfide crosslinks has been proposed as one of the misfolding pathways occurring in mutant SOD1; however, the pathological relevance of such oligomerization in the SOD1-ALS cases still remains obscure. We prepared antibodies exclusively recognizing the SOD1 oligomers cross-linked via disulfide bonds in vitro. By using those antibodies, immunohistochemical examination and ELISA were mainly performed on the tissue samples of transgenic mice expressing mutant SOD1 proteins and also of human SOD1-ALS cases. We showed the recognition specificity of our antibodies exclusively toward the disulfide-crosslinked SOD1 oligomers by ELISA using various forms of purified SOD1 proteins in conformationally distinct states in vitro. Furthermore, the epitope of those antibodies was buried and inaccessible in the natively folded structure of SOD1. The antibodies were then found to specifically detect the pathological SOD1 species in the spinal motor neurons of the SOD1-ALS patients as well as the transgenic model mice. Our findings here suggest that the SOD1 oligomerization through the disulfide-crosslinking associates with exposure of the SOD1 structural interior and is a pathological process occurring in the SOD1-ALS cases. The online version of this article (doi:10.1186/s13024-016-0145-9) contains supplementary material, which is available to authorized users.
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影响因子：
12.7
作者：
Forsberg K;Andersen PM;Marklund SL;Brännström T
通讯作者：
Brännström T
影响因子：
1.5
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Furukawa Y
通讯作者：
Furukawa Y
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Marklund, SL
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通讯作者：
Al-Chalabi, Ammar
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25
作者：
Kang, Shin H.;Li, Ying;Fukaya, Masahiro;Lorenzini, Ileana;Cleveland, Don W.;Ostrow, Lyle W.;Rothstein, Jeffrey D.;Bergles, Dwight E.
通讯作者：
Bergles, Dwight E.