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用于在DNA支架上组装感兴趣蛋白质的DNA结合衔接子。

DNA binding adaptors to assemble proteins of interest on DNA scaffold.

作者信息

Nakata Eiji, Dinh Huyen, Nguyen Thang Minh, Morii Takashi

机构信息

Institute of Advanced Energy, Kyoto University, Uji, Kyoto, Japan.

Institute of Advanced Energy, Kyoto University, Uji, Kyoto, Japan.

出版信息

Methods Enzymol. 2019;617:287-322. doi: 10.1016/bs.mie.2018.12.014. Epub 2019 Feb 13.

Abstract

DNA nanostructures serve as the ideal scaffolds to assemble materials of interest. Among these, proteins are of particularly interesting class of molecules to assemble because of their huge functional variability. Sequence-specific DNA binding proteins have been applied as adaptors to stably locate the fused proteins at defined positions of DNA scaffold in high loading yields. The strategy allows to control the number of enzyme molecules and to maintain the catalytic activity. By fusing a chemoselective self-ligating protein tag to the DNA binding protein, the modular adaptors formed covalent bonds at respective sequences on DNA scaffold with fast reaction kinetics. Application of a set of orthogonal modular adaptors enables spatial organization of multiple types of enzymes.

摘要

DNA纳米结构是组装目标材料的理想支架。其中,蛋白质因其巨大的功能多样性而成为特别有趣的一类可组装分子。序列特异性DNA结合蛋白已被用作衔接子,以高负载率将融合蛋白稳定定位在DNA支架的特定位置。该策略能够控制酶分子的数量并维持催化活性。通过将化学选择性自连接蛋白标签与DNA结合蛋白融合,模块化衔接子以快速的反应动力学在DNA支架上的相应序列处形成共价键。应用一组正交模块化衔接子能够实现多种类型酶的空间组织。

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