Institute for Biological Interfaces (IBG-1), Karlsruhe Institute of Technology (KIT), 76344, Eggenstein-Leopoldshafen, Germany.
School of Engineering, University of Applied Sciences Upper Austria, Wels, 4600, Austria.
Small. 2022 Sep;18(35):e2202704. doi: 10.1002/smll.202202704. Epub 2022 Aug 7.
News from an old acquaintance: The streptavidin (STV)-biotin binding system is frequently used for the decoration of DNA origami nanostructures (DON) to study biological systems. Here, a surprisingly high dynamic of the STV/DON interaction is reported, which is affected by the structure of the DNA linker system. Analysis of different mono- or bi-dentate linker architectures on DON with a novel high-speed atomic force microscope (HS-AFM) enabling acquisition times as short as 50 ms per frame gave detailed insights into the dynamics of the DON/STV interaction, revealing dwell times in the sub-100 millisecond range. The linker systems are also used to present biotinylated epidermal growth factor on DON to study the activation of the epidermal growth factor receptor signaling cascade in HeLa cells. The studies confirm that cellular activation correlated with the binding properties of linker-specific STV/DON interactions observed by HS-AFM. This work sheds more light on the commonly used STV/DON system and will help to further standardize the use of DNA nanostructures for the study of biological processes.
链霉亲和素（STV）-生物素结合系统常用于 DNA 折纸纳米结构（DON）的修饰，以研究生物系统。在这里，报道了 STV/DON 相互作用令人惊讶的高动态性，这受到 DNA 连接子系统结构的影响。使用一种新型高速原子力显微镜（HS-AFM）分析具有不同单齿或双齿连接子结构的 DON，该显微镜能够在每帧 50 毫秒的短时间内进行采集，从而深入了解 DON/STV 相互作用的动力学，揭示了亚 100 毫秒的停留时间。这些连接子系统还用于在 DON 上展示生物素化表皮生长因子，以研究表皮生长因子受体信号级联在 HeLa 细胞中的激活。研究证实，细胞激活与 HS-AFM 观察到的具有连接子特异性的 STV/DON 相互作用的结合特性相关。这项工作进一步揭示了常用的 STV/DON 系统，并将有助于进一步规范 DNA 纳米结构在生物过程研究中的应用。
