Herein, we report the biosynthesis of protein heterocatenanes using a programmed sequence of multiple post‐translational processing events including intramolecular chain entanglement, in situ backbone cleavage, and spontaneous cyclization. The approach is general, autonomous, and can obviate the need for any additional enzymes. The catenane topology was convincingly proven using a combination of SDS‐PAGE, LC‐MS, size exclusion chromatography, controlled proteolytic digestion, and protein crystallography. The X‐ray crystal structure clearly shows two mechanically interlocked protein rings with intact folded domains. It opens new avenues in the nascent field of protein‐topology engineering.

中文翻译：

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设计的蛋白质杂十八烯的细胞合成和X射线晶体结构。

在本文中，我们报告了使用多种翻译后加工事件（包括分子内链缠结，原位主链裂解和自发环化）的程序化序列进行蛋白质异儿烷的生物合成。该方法是通用的，自主的，并且可以消除对任何其他酶的需求。使用SDS-PAGE，LC-MS，尺寸排阻色谱，可控蛋白水解消化和蛋白质晶体学相结合，可证明链环烷拓扑结构令人信服。X射线晶体结构清楚地显示了两个具有完整折叠域的机械互锁的蛋白环。它为蛋白质拓扑结构工程的新生领域开辟了新途径。