Taninaka Atsushi, Takeuchi Osamu, Shigekawa Hidemi
Institute of Applied Physics, CREST-JST, University of Tsukuba/1-1-1 Tennodai, Tsukuba 305-8573, Japan; E-Mails:
Int J Mol Sci. 2010 May 13;11(5):2134-51. doi: 10.3390/ijms11052134.
To understand and design molecular functions on the basis of molecular recognition processes, the microscopic probing of the energy landscapes of individual interactions in a molecular complex and their dependence on the surrounding conditions is of great importance. Dynamic force spectroscopy (DFS) is a technique that enables us to study the interaction between molecules at the single-molecule level. However, the obtained results differ among previous studies, which is considered to be caused by the differences in the measurement conditions. We have developed an atomic force microscopy technique that enables the precise analysis of molecular interactions on the basis of DFS. After verifying the performance of this technique, we carried out measurements to determine the landscapes of streptavidin-biotin interactions. The obtained results showed good agreement with theoretical predictions. Lifetimes were also well analyzed. Using a combination of cross-linkers and the atomic force microscope that we developed, site-selective measurement was carried out, and the steps involved in bonding due to microscopic interactions are discussed using the results obtained by site-selective analysis.
为了基于分子识别过程理解和设计分子功能，对分子复合物中单个相互作用的能量景观及其对周围条件的依赖性进行微观探测至关重要。动态力谱（DFS）是一种使我们能够在单分子水平上研究分子间相互作用的技术。然而，先前研究所得结果存在差异，这被认为是由测量条件的差异所致。我们开发了一种基于DFS能够精确分析分子相互作用的原子力显微镜技术。在验证了该技术的性能之后，我们进行了测量以确定链霉亲和素 - 生物素相互作用的景观。所得结果与理论预测吻合良好。寿命也得到了很好的分析。使用我们开发的交联剂和原子力显微镜的组合进行了位点选择性测量，并利用位点选择性分析所得结果讨论了微观相互作用导致的键合步骤。
