Single-molecule bonds characterized by solid-state nanopore force spectroscopy.
Single-molecule bonds characterized by solid-state nanopore force spectroscopy.
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DOI：
10.1021/nn900713a
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发表时间：
2009-10-27
期刊：
影响因子：
17.1
通讯作者：
Marziali A
中科院分区：
文献类型：
--
作者：
Tabard-Cossa V;Wiggin M;Trivedi D;Jetha NN;Dwyer JR;Marziali A
弱分子相互作用驱动生命系统核心的过程，如酶-底物相互作用、受体-配体结合和核酸复制。单分子力谱是揭示非共价键的分子尺度能量景观的显着工具，通过直接对单个分子复合物施加机械力，并跟踪其作为时间和施加力的函数的存活。原则上，通过直接表征探针和靶分子之间的键合强度，力谱法也可用于高度特异性的分子识别测定。然而，传统的力谱技术的复杂性和低吞吐量使得这样的生物传感应用不切实际。在这里，我们展示了一种简单的单分子方法，适用于生物物理研究和分子识别测定，其中~3 nm的氮化硅纳米孔用于确定生物素-中性亲和素复合物的键寿命光谱。成千上万的单个分子复合物在恒定施加的力（从400 mV到900 mV）下在固态纳米孔中被捕获和解离，使我们能够提取控制相互作用的能垒的位置，映射在Δx = 0.5 nm处。这些结果突出了固态纳米孔检测和表征分子间相互作用的能力，并展示了如何将其应用于快速，高度特异性的分子检测分析。
Weak molecular interactions drive processes at the core of living systems, such as enzyme-substrate interactions, receptor-ligand binding, and nucleic acid replication. Single molecule force spectroscopy is a remarkable tool for revealing molecular scale energy landscapes of noncovalent bonds, by exerting a mechanical force directly on an individual molecular complex, and tracking its survival as a function of time and applied force. In principle, force spectroscopy methods can also be used for highly specific molecular recognition assays, by directly characterizing the strength of bonds between probe and target molecules. However, complexity and low throughput of conventional force spectroscopy techniques renders such biosensing applications impractical. Here we demonstrate a straightforward single molecule approach, suitable for both biophysical studies and molecular recognition assays, in which a ~3 nm silicon nitride nanopore is used to determine the bond lifetime spectrum of the biotin-neutravidin complex. Thousands of individual molecular complexes are captured and dissociated in the solid-state nanopore under constant applied forces, ranging from 400 mV to 900 mV, allowing us to extract the location of the energy barrier that governs the interaction, mapped at Δx ≈ 0.5 nm. These results highlight the capacity of a solid-state nanopore to detect and characterize inter-molecular interactions, and demonstrate how this could be applied to rapid, highly specific molecular detection assays.
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DOI：
10.1073/pnas.0806085105
发表时间：
2008-10-14
影响因子：
11.1
作者：
Dudko, Olga K.;Hummer, Gerhard;Szabo, Attila
通讯作者：
Szabo, Attila
影响因子：
56.9
作者：
Grubmuller, H;Heymann, B;Tavan, P
通讯作者：
Tavan, P
影响因子：
3.4
作者：
Wiggin, Matthew;Tropini, Carolina;Marziali, Andre
通讯作者：
Marziali, Andre
影响因子：
19.6
作者：
Keyser, Ulrich F.;Koeleman, Bernard N.;Dekker, Cees
通讯作者：
Dekker, Cees
影响因子：
3.4
作者：
Evans, E;Ritchie, K
通讯作者：
Ritchie, K