Intracellular targeting and clearance of oligomeric alpha-synuclein alleviates toxicity in mammalian cells
Intracellular targeting and clearance of oligomeric alpha-synuclein alleviates toxicity in mammalian cells
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DOI：
10.1016/j.neulet.2009.04.046
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发表时间：
2009-07-31
影响因子：
2.5
通讯作者：
Sierks, Michael R.
中科院分区：
文献类型：
--
作者：
Yuan, Bin;Sierks, Michael R.
有毒蛋白质聚集体的细胞内清除代表了治疗帕金森病和亨廷顿舞蹈病等蛋白质错误折叠疾病的一种有前景的治疗方法。细胞内表达的抗体片段或胞内抗体可用于结合特定的细胞内靶标。添加非传统的分泌信号序列使得胞内抗体首先在细胞内结合其靶标，然后将结合的靶标穿梭穿过细胞膜，将其从细胞中分泌出来。我们在过度表达 a-syn 的哺乳动物细胞模型中，在细胞内表达了两种不同的单链抗体 (scFv) 片段，靶向单体或寡聚 α-突触核蛋白 (a-syn)。研究了每个体内抗体的两种版本，一种具有非传统分泌信号，另一种不具有非传统分泌信号。靶向单体a-syn的scFv很少或没有降低由a-syn过度表达引起的毒性，然而寡聚a-syn的结合和分泌完全降低了毒性。因此，非传统的体内分泌代表了一种靶向和清除多种有害细胞内物质的有效方法。 (C) 2009 Elsevier Ireland Ltd. 保留所有权利。
Intracellular clearance of toxic protein aggregates represents a promising therapeutic approach to treat protein-misfolding diseases such as Parkinson's and Huntington's diseases. Intracelluarly expressed antibody fragments or intrabodies can be used to bind specific intracellular targets. Addition of a non-traditional secretion signal sequence enables the intrabody to first bind its target inside the cell and then shuttle the bound target through the cell membrane, secreting it from the cell. We intracellularly expressed two different single chain antibody (scFv) fragments targeting either monomeric or oligomeric alpha-synuclein (a-syn), in a mammalian cell model that overexpresses a-syn. Two versions of each intrabody were studied, one with and one without the non-traditional secretion signal. The scFv targeting monomeric a-syn provided little or no reduction in toxicity induced by overexpression of a-syn, however binding and secretion of oligomeric a-syn totally reduced toxicity. Non-traditional intrabody secretion therefore represents an effective method to target and clear a variety of harmful intracellular species. (C) 2009 Elsevier Ireland Ltd. All rights reserved.