An in vivo platform for identifying inhibitors of protein aggregation.
An in vivo platform for identifying inhibitors of protein aggregation.
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DOI：
10.1038/nchembio.1988
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发表时间：
2016-02
影响因子：
14.8
通讯作者：
Radford SE
中科院分区：
文献类型：
--
作者：
Saunders JC;Young LM;Mahood RA;Jackson MP;Revill CH;Foster RJ;Smith DA;Ashcroft AE;Brockwell DJ;Radford SE
蛋白质聚集是一系列人类疾病的基础，但迄今为止只有一种小分子治疗剂被成功开发。在这里，我们介绍了一种体内系统，基于β-内酰胺酶三方融合构建体，能够识别大肠杆菌周质中的易聚集序列和防止其异常自组装的抑制剂。我们使用一系列蛋白质证明了该系统的功能，从小的非结构化肽（胰岛淀粉样多肽和淀粉样β蛋白）到较大的折叠免疫球蛋白结构域。以48孔形式配置，分裂β-内酰胺酶传感器容易区分易聚集序列和可溶性序列。在109种化合物的存在下进行测定使得抑制的等级排序成为可能，并揭示了IAPP聚集的新抑制剂。该平台可以应用于淀粉样蛋白生成和其他聚集倾向系统，与序列或大小无关，并且可以鉴定能够改善或抑制蛋白质聚集的小分子或其他因子。
Protein aggregation underlies an array of human diseases, yet only one small molecule therapeutic has been successfully developed to date. Here, we introduce an in vivo system, based on a β-lactamase tripartite fusion construct, capable of identifying aggregation-prone sequences in the periplasm of Escherichia coli and inhibitors that prevent their aberrant self-assembly. We demonstrate the power of the system using a range of proteins, from small unstructured peptides (islet amyloid polypeptide and amyloid β) to larger, folded immunoglobulin domains. Configured in a 48-well format, the split β-lactamase sensor readily differentiates between aggregation-prone and soluble sequences. Performing the assay in the presence of 109 compounds enabled a rank ordering of inhibition and revealed a new inhibitor of IAPP aggregation. This platform can be applied to both amyloidogenic and other aggregation-prone systems, independent of sequence or size, and can identify small molecules or other factors able to ameliorate or inhibit protein aggregation.