Imanishi Miki, Negi Shigeru, Sugiura Yukio
Institute for Chemical Research, Kyoto University, Kyoto, Japan.
Methods Mol Biol. 2010;649:337-49. doi: 10.1007/978-1-60761-753-2_21.
The design of functional proteins is one of the most challenging areas of protein research. We have constructed zinc finger peptides with metal-dependent hydrolytic abilities by mutating the zinc ligands in classical zinc fingers, without the need to add a FokI or other DNA cleavage domain. The designed peptides acquired DNA cleavage ability successfully, retaining the proper zinc finger folding and DNA targeting ability. We have also succeeded in site-specific DNA cleavage in the presence of cerium ions by introducing a lanthanide ion-binding loop as a linker of zinc finger motifs.
功能性蛋白质的设计是蛋白质研究中最具挑战性的领域之一。我们通过突变经典锌指中的锌配体，构建了具有金属依赖性水解能力的锌指肽，无需添加FokI或其他DNA切割结构域。所设计的肽成功获得了DNA切割能力，同时保留了适当的锌指折叠和DNA靶向能力。我们还通过引入镧系离子结合环作为锌指基序的连接体，在铈离子存在的情况下成功实现了位点特异性DNA切割。
