Department of Molecular Microbiology and Biotechnology, Tel Aviv University, Tel Aviv, Israel.
PLoS One. 2010 Nov 10;5(11):e13863. doi: 10.1371/journal.pone.0013863.
The intracellular oligomerization of α-synuclein is associated with Parkinson's disease and appears to be an important target for disease-modifying treatment. Yet, to date, there is no specific inhibitor for this aggregation process. Using unbiased systematic peptide array analysis, we identified molecular interaction domains within the β-synuclein polypeptide that specifically binds α-synuclein. Adding such peptide fragments to α-synuclein significantly reduced both amyloid fibrils and soluble oligomer formation in vitro. A retro-inverso analogue of the best peptide inhibitor was designed to develop the identified molecular recognition module into a drug candidate. While this peptide shows indistinguishable activity as compared to the native peptide, it is stable in mouse serum and penetrates α-synuclein over-expressing cells. The interaction interface between the D-amino acid peptide and α-synuclein was mapped by Nuclear Magnetic Resonance spectroscopy. Finally, administering the retro-inverso peptide to a Drosophila model expressing mutant A53T α-synuclein in the nervous system, resulted in a significant recovery of the behavioral abnormalities of the treated flies and in a significant reduction in α-synuclein accumulation in the brains of the flies. The engineered retro-inverso peptide can serve as a lead for developing a novel class of therapeutic agents to treat Parkinson's disease.
α-突触核蛋白的细胞内寡聚化与帕金森病有关，似乎是一种针对疾病修饰治疗的重要靶点。然而，迄今为止，还没有针对这种聚集过程的特异性抑制剂。通过无偏系统肽阵列分析，我们确定了β-突触核蛋白多肽内特异性结合α-突触核蛋白的分子相互作用域。向α-突触核蛋白中添加这些肽片段可显著减少体外淀粉样纤维和可溶性寡聚物的形成。设计了一种最佳肽抑制剂的反向非对映异构体，以将鉴定出的分子识别模块开发成候选药物。虽然该肽与天然肽相比具有相同的活性，但它在小鼠血清中稳定且可穿透过表达α-突触核蛋白的细胞。通过核磁共振波谱确定了 D-氨基酸肽与 α-突触核蛋白之间的相互作用界面。最后，将反向非对映肽施用于在神经系统中表达突变 A53T α-突触核蛋白的果蝇模型中，导致处理过的果蝇的行为异常得到显著恢复，并且果蝇大脑中α-突触核蛋白的积累显著减少。该工程反向非对映肽可作为开发治疗帕金森病的新型治疗剂的先导。
