Tsuji-Ueno Sachika, Komatsu Masayuki, Iguchi Kakeru, Takahashi Masahiro, Yoshino Syuhei, Suzuki Miho, Nemoto Naoto, Nishigaki Koichi
Department of Functional Materials Science, Graduate School of Science and Engineering, Saitama University, Saitama 338-8570, Japan.
Protein Pept Lett. 2011 Jun;18(6):642-50. doi: 10.2174/092986611795222678.
Recent studies have been supporting that the generation of Aβ42 oligomers is responsible for Alzheimer's disease. Therefore, those peptides which bind to Aβ42 are scientifically interesting and can be possible candidates for the diagnosis and therapy of Alzheimer's disease. A systemic in vitro evolution, developed recently and the designated progressive library method (PLM), was applied to obtain Ab42-binding aptamers peptides. As a result, high affinity peptide aptamers made of 8 or 9 amino acids could be identified by this approach, endorsing the methodological effectiveness. Namely, the selection products from the secondary library of diversified peptides, which was constructed based on the information obtained from the primary library selection, were confirmed to be superior to those selected from the primary library as had been reported previously. The affinities of those peptides measured by SPR (surface plasmon resonance) were comparable to or higher than that of those peptides so far reported (K(d) of 10⁻⁷). The other peptides selected were confirmed of their binding by a novel mode of gel shift assay (fluorescence enhancement caused by the binding). Thus, novel Aβ42-binding peptides with high affinity were provided for the future Alzheimer's disease study. The demonstration of the effectiveness of the systemic in vitro evolution of PLM is very encouraging for the study of identifying novel functional peptides.
最近的研究支持β淀粉样蛋白42（Aβ42）寡聚体的产生是阿尔茨海默病的病因。因此，那些与Aβ42结合的肽在科学上具有重要意义，并且可能成为阿尔茨海默病诊断和治疗的候选药物。最近开发的一种系统体外进化方法，即指定的渐进文库法（PLM），被用于获得与Aβ42结合的适体肽。结果，通过这种方法可以鉴定出由8个或9个氨基酸组成的高亲和力肽适体，证明了该方法的有效性。也就是说，基于从一级文库筛选获得的信息构建的多样化肽二级文库的筛选产物，被证实优于先前报道的从一级文库中筛选出的产物。通过表面等离子体共振（SPR）测量的这些肽的亲和力与迄今为止报道的那些肽相当或更高（解离常数K(d)为10⁻⁷）。通过一种新的凝胶迁移试验模式（结合引起的荧光增强）证实了其他所选肽的结合。因此，为未来的阿尔茨海默病研究提供了具有高亲和力的新型Aβ42结合肽。PLM系统体外进化有效性的证明对于鉴定新型功能肽的研究非常鼓舞人心。
