van der Sleen Lyan M, Tych Katarzyna M
Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, 9747 AG Groningen, The Netherlands.
Nanomaterials (Basel). 2021 Sep 17;11(9):2424. doi: 10.3390/nano11092424.
The mechanical properties of proteins can be studied with single molecule force spectroscopy (SMFS) using optical tweezers, atomic force microscopy and magnetic tweezers. It is common to utilize a flexible linker between the protein and trapped probe to exclude short-range interactions in SMFS experiments. One of the most prevalent linkers is DNA due to its well-defined properties, although attachment strategies between the DNA linker and protein or probe may vary. We will therefore provide a general overview of the currently existing non-covalent and covalent bioconjugation strategies to site-specifically conjugate DNA-linkers to the protein of interest. In the search for a standardized conjugation strategy, considerations include their mechanical properties in the context of SMFS, feasibility of site-directed labeling, labeling efficiency, and costs.
蛋白质的机械性能可以通过使用光镊、原子力显微镜和磁镊的单分子力谱（SMFS）进行研究。在SMFS实验中，通常在蛋白质和捕获探针之间使用柔性接头来排除短程相互作用。由于DNA具有明确的特性，因此它是最常用的接头之一，尽管DNA接头与蛋白质或探针之间的连接策略可能有所不同。因此，我们将概述目前现有的非共价和共价生物共轭策略，以便将DNA接头位点特异性地共轭到感兴趣的蛋白质上。在寻找标准化的共轭策略时，需要考虑的因素包括它们在SMFS背景下的机械性能、定点标记的可行性、标记效率和成本。
