Rational design of amyloid β peptide–binding proteins: Pseudo‐Aβ β‐sheet surface presented in green fluorescent protein binds tightly and preferentially to structured Aβ
Rational design of amyloid β peptide–binding proteins: Pseudo‐Aβ β‐sheet surface presented in green fluorescent protein binds tightly and preferentially to structured Aβ
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淀粉样蛋白 β 肽结合蛋白的合理设计：绿色荧光蛋白中存在的伪 Aβ β 片层表面优先与结构化 Aβ 紧密结合
DOI：
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
H. Mihara
中科院分区：
文献类型：
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作者：
Tsuyoshi Takahashi;K. Ohta;H. Mihara
一些神经退行性疾病，如阿尔茨海默病(AD)和帕金森病，是由蛋白质错误折叠引起的。在阿尔茨海默病中，淀粉样蛋白β肽(Aβ)被认为是一种有毒物质，它通过自组装形成涉及可溶性寡聚中间体和淀粉样纤维的各种聚集体。在这里，我们设计了几种含有伪Aββ-Sheet表面的绿色荧光蛋白突变体，并评估了它们与Aβ结合和抑制Aβ齐聚的能力。两个绿色荧光蛋白突变体P13H和AP93Q与Aβ紧密结合，Kd=2 6 0 nM和Kd=42 0 nM。细胞存活率实验表明，P13H和AP93Q能有效抑制毒性Aβ寡聚体的产生。通过结合P13H和AP93Q突变，设计了一个由四条Aβ衍生序列组成的超变异体SFAB4，它与Aβ(Kd=100nM)的结合比只有两条伪Aβ链的SFAB4更紧密。SFAB4蛋白比非结构单体和成熟的淀粉样纤维更能识别Aβ的可溶性低聚中间体。因此，将伪Aββ-Sheet结构嵌入到以β-Barrel结构排列的蛋白质表面的设计策略对于构建能够与Aβ紧密结合并抑制其聚集的分子是有用的。这一策略可能为AD的诊断和治疗的发展提供参考。蛋白质2010年。©2009威利-利斯，Inc.
Some neurodegenerative diseases such as Alzheimer disease (AD) and Parkinson disease are caused by protein misfolding. In AD, amyloid β‐peptide (Aβ) is thought to be a toxic agent by self‐assembling into a variety of aggregates involving soluble oligomeric intermediates and amyloid fibrils. Here, we have designed several green fluorescent protein (GFP) variants that contain pseudo‐Aβ β‐sheet surfaces and evaluated their abilities to bind to Aβ and inhibit Aβ oligomerization. Two GFP variants P13H and AP93Q bound tightly to Aβ, Kd = 260 nM and Kd = 420 nM, respectively. Moreover, P13H and AP93Q were capable of efficiently suppressing the generation of toxic Aβ oligomers as shown by a cell viability assay. By combining the P13H and AP93Q mutations, a super variant SFAB4 comprising four strands of Aβ‐derived sequences was designed and bound more tightly to Aβ (Kd = 100 nM) than those having only two pseudo‐Aβ strands. The SFAB4 protein preferentially recognized the soluble oligomeric intermediates of Aβ more than both unstructured monomer and mature amyloid fibrils. Thus, the design strategy for embedding pseudo‐Aβ β‐sheet structures onto a protein surface arranged in the β‐barrel structure is useful to construct molecules capable of binding tightly to Aβ and inhibiting its aggregation. This strategy may provide implication for the diagnostic and therapeutic development in the treatment of AD. Proteins 2010. © 2009 Wiley‐Liss, Inc.
DOI：
10.1006/bbrc.1996.1413
发表时间：
1996-09-24
影响因子：
3.1
作者：
Soto, C;Kindy, MS;Frangione, B
通讯作者：
Frangione, B