Tissue damage in the amyloidoses: Transthyretin monomers and nonnative oligomers are the major cytotoxic species in tissue culture
Tissue damage in the amyloidoses: Transthyretin monomers and nonnative oligomers are the major cytotoxic species in tissue culture
复制标题
DOI：
10.1073/pnas.0400062101
复制
发表时间：
2004-03-02
影响因子：
11.1
通讯作者：
Buxbaum, JN
中科院分区：
文献类型：
--
作者：
Reixach, N;Deechongkit, S;Buxbaum, JN
转甲状腺素蛋白 (TTR) 淀粉样变性是一种人类疾病，其中错误折叠的 TTR 蛋白在组织中聚集，随后导致内脏、外周和自主神经功能障碍。最近的报告强调了寡聚中间体作为各种形式的淀粉样蛋白生成中主要细胞毒性物质的重要性。我们研究了野生型和变体 TTR 蛋白的几种四级结构状态对神经谱系细胞的细胞毒性作用。 TTR 淀粉样原纤维和>100 kDa 的可溶性聚集体没有毒性。在细胞测定条件下孵育 TTR，并通过尺寸排阻色谱法和 SDS/PAGE 进行分析表明，单体 TTR 或相对较小、快速形成的最大尺寸为 6 个亚基的聚集体是主要的细胞毒性物质。稳定天然四聚体状态的小分子被证明可以防止毒性。这些研究与错误折叠的 TTR 单体快速聚集形成瞬时低分子质量组装体的模型一致。
The transthyretin (TTR) amyloidoses are human diseases in which the misfolded TTR protein aggregates in tissues with subsequent visceral, peripheral, and autonomic nerve dysfunction. Recent reports have stressed the importance of oligomeric intermediates as major cytotoxic species in various forms of amyloidogenesis. We have examined the cytotoxic effects of several quaternary structural states of wild-type and variant TTR proteins on cells of neural lineage. TTR amyloid fibrils and soluble aggregates >100 kDa were not toxic. Incubation of TTR under the conditions of the cell assay and analysis by size-exclusion chromatography and SDS/PAGE reveal that monomeric TTR or relatively small, rapidly formed aggregates of a maximum size of six subunits were the major cytotoxic species. Small molecules that stabilize the native tetrameric state were shown to prevent toxicity. The studies are consistent with a model in which the misfolded TTR monomer rapidly aggregates to form transient low molecular mass assemblies (