In-cell NMR reveals potential precursor of toxic species from SOD1 fALS mutants
In-cell NMR reveals potential precursor of toxic species from SOD1 fALS mutants
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DOI：
10.1038/ncomms6502
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发表时间：
2014-11-01
影响因子：
16.6
通讯作者：
Banci, Lucia
中科院分区：
文献类型：
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作者：
Luchinat, Enrico;Barbieri, Letizia;Banci, Lucia
超氧化物歧化酶1（SOD1）基因突变与肌萎缩侧索硬化症（fALS）的家族性病例有关。在这里，我们利用细胞内NMR表征一系列fALS连接的SOD1突变体在人类细胞中的蛋白质折叠和成熟，并在分子水平上深入了解它们在细胞中的行为。通过改变细胞中金属离子的可用性，以及通过共表达SOD 1的铜分子伴侣hCCS，研究了各种突变对SOD 1成熟的影响。我们观察到大多数突变体的发生的非结构化的SOD 1物种，无法结合锌。这种物质可能是与fALS相关的潜在毒性低聚物物质的常见前体。在铜的存在下，hCCS的共表达恢复了SOD1突变体的正确成熟，并防止了非结构化物种的形成，证实了hCCS也充当分子伴侣。
Mutations in the superoxide dismutase 1 (SOD1) gene are related to familial cases of amyotrophic lateral sclerosis (fALS). Here we exploit in-cell NMR to characterize the protein folding and maturation of a series of fALS-linked SOD1 mutants in human cells and to obtain insight into their behaviour in the cellular context, at the molecular level. The effect of various mutations on SOD1 maturation are investigated by changing the availability of metal ions in the cells, and by coexpressing the copper chaperone for SOD1, hCCS. We observe for most of the mutants the occurrence of an unstructured SOD1 species, unable to bind zinc. This species may be a common precursor of potentially toxic oligomeric species, that are associated with fALS. Coexpression of hCCS in the presence of copper restores the correct maturation of the SOD1 mutants and prevents the formation of the unstructured species, confirming that hCCS also acts as a molecular chaperone.