工程化结构域交换作为蛋白质功能的开/关开关
Engineered Domain Swapping as an On/Off Switch for Protein Function.
作者信息
Ha Jeung-Hoi, Karchin Joshua M, Walker-Kopp Nancy, Castañeda Carlos A, Loh Stewart N
机构信息
Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University, 750 East Adams Street, Syracuse, NY 13210, USA.
Departments of Biology and Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244, USA.
出版信息
Chem Biol. 2015 Oct 22;22(10):1384-93. doi: 10.1016/j.chembiol.2015.09.007.
Domain swapping occurs when identical proteins exchange segments in reciprocal fashion. Natural swapping mechanisms remain poorly understood, and engineered swapping has the potential for creating self-assembling biomaterials that encode for emergent functions. We demonstrate that induced swapping can be used to regulate the function of a target protein. Swapping is triggered by inserting a "lever" protein (ubiquitin) into one of four loops of the ribose binding protein (RBP) target. The lever splits the target, forcing RBP to refold in trans to generate swapped oligomers. Identical RBP-ubiquitin fusions form homo-swapped complexes with the ubiquitin domain acting as the hinge. Surprisingly, some pairs of non-identical fusions swap more efficiently with each other than they do with themselves. Nuclear magnetic resonance experiments reveal that the hinge of these hetero-swapped complexes maps to a region of RBP distant from both ubiquitins. This design is expected to be applicable to other proteins to convert them into functional switches.
当相同的蛋白质以相互交换的方式交换片段时，就会发生结构域交换。天然的交换机制仍知之甚少，而工程化交换有潜力创造出编码新功能的自组装生物材料。我们证明诱导交换可用于调节靶蛋白的功能。通过将“杠杆”蛋白（泛素）插入核糖结合蛋白（RBP）靶标的四个环之一中来触发交换。杠杆将靶标分开，迫使RBP反式折叠以产生交换的寡聚体。相同的RBP-泛素融合体形成同型交换复合物，其中泛素结构域充当铰链。令人惊讶的是，一些非相同融合体对彼此之间的交换比它们自身之间的交换更有效。核磁共振实验表明，这些异源交换复合物的铰链映射到RBP中远离两个泛素的区域。预计这种设计适用于其他蛋白质，将它们转化为功能开关。
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