Cellular Synthesis of Protein Catenanes
Cellular Synthesis of Protein Catenanes
复制标题
蛋白质索烷的细胞合成。
DOI：
10.1002/anie.201511640
复制
发表时间：
2016-03-01
影响因子：
16.6
通讯作者：
Zhang, Wen-Bin
中科院分区：
文献类型：
--
作者：
Wang, Xiao-Wei;Zhang, Wen-Bin
细胞直接生产拓扑结构复杂的蛋白质是超分子化学和蛋白质工程领域的研究热点。我们描述了通过使用P53二聚化结构域来指导两个蛋白链的交织和SpyTag-Spycatcher化学来高效环化来首次在细胞内合成蛋白质连环。SDS-PAGE、SEC和部分消化实验明确地证明了Catenane的拓扑结构，并且与单体控制突变体相比，它对胰酶的消化具有更高的稳定性。由蛋白质折叠和基因编码的蛋白质化学实现的组装-反应协同作用为在 vivo中创建机械连锁的复杂蛋白质拓扑提供了一种方便而强大的方法。
Direct cellular production of topologically complex proteins is of great interest both in supramolecular chemistry and protein engineering. We describe the first cellular synthesis of protein catenanes through the use of the p53 dimerization domain to guide the intertwining of two protein chains and SpyTag-SpyCatcher chemistry for efficient cyclization. The catenane topology was unambiguously proven by SDS-PAGE, SEC, and partial digestion experiments and was shown to confer enhanced stability toward trypsin digestion relative to monomeric control mutants. The assembly-reaction synergy enabled by protein folding and genetically encoded protein chemistry offers a convenient yet powerful approach for creating mechanically interlocked, complex protein topologies in vivo.