Single Molecule Translocation through Biological Nanopores with Optical Tweezers

在拟议的研究项目中，我们计划使用光镊力力学来研究和量化单个生物聚合物通过生物纳米孔（biopores）的移位机制和动力学（亚）分子分辨率。这种单分子技术允许测量有效的分子力和弹性，机械噪声谱，易位动力学，相关的能量景观以及相关的纳米流体特性，而无需分子标记和扩增。我们将研究如何链状大分子，如核酸，多糖，以及折叠和未折叠的蛋白质将通过生物孔（孔蛋白），功能集成在支持的生物膜和仿生脂质杂交体，结合单分子力谱与纳米流体的多个方面。由于单分子易位是由热非平衡，快速热波动和大分子随机性，我们的目标是一个快速的力和噪声分析与随机模拟的实验结果的连贯描述（合作：P. Reimann）。我们的目标是深入的基本机制的见解，通过一般的生物孔的分子易位，以及纳米生物技术的应用。后者依赖于在通过生物纳米孔易位期间识别特定的亚分子力特征，从而能够推断易位大分子的结构信息（例如DNA的甲基化模式）。

In the proposed research project we plan to use optical tweezers force mechanics to investigate and quantify the translocation mechanisms and dynamics of single biopolymers through biological nanopores (biopores) with (sub)molecular resolution. This single molecule technology allows to measure effective molecular forces and elasticities, mechanical noise spectra, translocation dynamics, associated energy landscape as well as associated nanofluidic properties without molecule labelling and amplification. We will study how chain-like macromolecules like nucleic acids, polysaccharides, as well as folded and unfolded proteins will be threaded through biopores (porins) that are functionally integrated in supported biomembranes and biomimetic lipid-hybrids thereof, combining manifold aspects of single-molecule force spectroscopy with nanofluidics. Since single-molecule translocation is governed by thermal non-equilibrium, fast thermal fluctuations and macromolecular stochastics, we aim for a fast force and noise analysis together with a coherent description of the experimental results with stochastic simulations (Cooperation: P. Reimann). We aim for deep fundamental mechanistic insights into the molecule translocation through biopores in general, as well as nanobiotechnological applications thereof. The latter rely on the identification of specific, sub-molecular force signatures during translocation through biological nanopores, allowing to deduce structural information of the translocated macromolecule (e.g. the methylation pattern of DNA).