Single Molecule Translocation through Biological Nanopores with Optical Tweezers

使用光镊通过生物纳米孔进行单分子易位

• 批准号: 269589970
• 负责人: Professor Dr. Dario Anselmetti
• 金额: --
• 依托单位: Arbeitsgruppe Theorie der Kondensierten Materie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/269589970?language=en
• 关键词: Single; Molecule; Translocation; through; Biological

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).

在提出的研究项目中，我们计划使用光钳作用力机制来研究和量化单个生物聚合物通过具有(亚)分子分辨率的生物纳米孔(生物孔)的移位机制和动力学。这种单分子技术允许测量有效的分子力和弹性、机械噪声光谱、移位动力学、相关的能量景观以及相关的纳米流体性质，而无需分子标记和放大。我们将研究链状大分子，如核酸，多糖，以及折叠和未折叠的蛋白质将如何穿过生物孔(孔蛋白)，这些孔在功能上整合在支撑的生物膜及其仿生脂质杂交物中，将单分子力谱的多个方面与纳米流体相结合。由于单分子移位受热非平衡、快速热涨落和大分子随机性的控制，我们的目标是快速的力和噪声分析，并通过随机模拟对实验结果进行连贯的描述(合作：P.Reimann)。我们的目标是深入了解通过生物孔进行的分子移位以及纳米生物技术在其中的应用。后者依赖于在通过生物纳米孔的移位过程中识别特定的亚分子作用力信号，从而能够推断出移位的大分子的结构信息(例如DNA的甲基化模式)。

项目成果

Preparation of 2D Phospholipid and Copolymer Nanomembranes

二维磷脂和共聚物纳米膜的制备

• DOI: 10.1016/j.matpr.2017.09.172
• 期刊: Materials Today: Proceedings
• 作者: R. Hillmann;D. Gilzer;N. Biere;S. Knust;M. Viefhues;K. Toensing;T. Kottke;D. Anselmetti
• 通讯作者: D. Anselmetti

Measuring DNA Translocation Forces through MoS 2 -Nanopores with Optical Tweezers

• DOI: 10.1016/j.matpr.2017.09.182
• 发表时间: 2017
• 期刊: Materials Today: Proceedings
• 作者: Sebastian Knust;Dennis Kreft;Roland Hillmann;Andreas J Meyer;Martina Viefhues;P. Reimann;D. Anselmetti

Biphasic Formation of 2D Nanomembranes by Photopolymerization of Diacetylene Lipids as Revealed by Infrared Difference Spectroscopy.

• DOI: 10.1021/acs.langmuir.9b00660
• 发表时间: 2019-06
• 期刊: Langmuir : the ACS journal of surfaces and colloids
• 作者: Dominic Gilzer;Roland Hillmann;Lukas Goett-Zink;Jessica L. Klocke;Martina Viefhues;D. Anselmetti;T. Kottke

Characterization of Robust and Free-Standing 2D-Nanomembranes of UV-Polymerized Diacetylene Lipids.

紫外聚合二乙炔脂质的坚固且独立的二维纳米膜的表征

• DOI: 10.1021/acs.langmuir.7b03403
• 发表时间: 2018
• 期刊: Langmuir : the ACS journal of surfaces and colloids
• 作者: R. Hillmann;M. Viefhues;L. Goett-Zink;D. Gilzer;Th. Hellweg;A. Gölzhäuser;T. Kottke;D. Anselmetti
• 通讯作者: D. Anselmetti