Protein analysis by time-resolved measurements with an electro-switchable DNA chip.
Protein analysis by time-resolved measurements with an electro-switchable DNA chip.
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DOI：
10.1038/ncomms3099
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发表时间：
2013
影响因子：
16.6
通讯作者：
Rant, Ulrich
中科院分区：
文献类型：
--
作者：
Langer, Andreas;Hampel, Paul A.;Kaiser, Wolfgang;Knezevic, Jelena;Welte, Thomas;Villa, Valentina;Maruyama, Makiko;Svejda, Matej;Jaehner, Simone;Fischer, Frank;Strasser, Ralf;Rant, Ulrich
在固定相或流动相中进行测量是蛋白质分析的基本原理。虽然分子固定在固体载体上可以并行分析相互作用，但大小或形状等性质通常是从分子在外力影响下的流动性推断出来的。然而，由于这些原则是相互排斥的，蛋白质的全面表征通常涉及一个多步骤的工作流程。在这里，我们展示了如何通过动态驱动的纳米棒将蛋白质拴在表面来协调这些测量模式。在交变电场的作用下，短链DNA被用作捕获探针与目标蛋白结合。通过在纳米振幅上摆动蛋白质，并将它们的运动动力学与理论模型进行比较，可以以Angström精度量化蛋白质直径。三级蛋白质结构(折叠)和构象的变化很容易被检测到，甚至翻译后的修饰也可以通过时间分辨的分子动力学测量来揭示。蛋白质的综合生物分析通常需要多步骤的表面和流动相测量。在这里，作者使用带有动态驱动的纳米棒的芯片--可以在电场中切换的DNA链--来获得芯片上蛋白质的动态测量。
Measurements in stationary or mobile phases are fundamental principles in protein analysis. Although the immobilization of molecules on solid supports allows for the parallel analysis of interactions, properties like size or shape are usually inferred from the molecular mobility under the influence of external forces. However, as these principles are mutually exclusive, a comprehensive characterization of proteins usually involves a multi-step workflow. Here we show how these measurement modalities can be reconciled by tethering proteins to a surface via dynamically actuated nanolevers. Short DNA strands, which are switched by alternating electric fields, are employed as capture probes to bind target proteins. By swaying the proteins over nanometre amplitudes and comparing their motional dynamics to a theoretical model, the protein diameter can be quantified with Angström accuracy. Alterations in the tertiary protein structure (folding) and conformational changes are readily detected, and even post-translational modifications are revealed by time-resolved molecular dynamics measurements. The comprehensive bioanalysis of proteins usually requires multi-step surface and mobile phase measurements. Here, the authors use chips functionalized with dynamically actuated nanolevers—DNA strands that can be switched in an electric field—to obtain motional dynamic measurements of proteins on a chip.
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影响因子：
7.4
作者：
Kussrow, Amanda;Enders, Carolyn S.;Bornhop, Darryl J.
通讯作者：
Bornhop, Darryl J.
影响因子：
15
作者：
Knezevic, Jelena;Langer, Andreas;Rant, Ulrich
通讯作者：
Rant, Ulrich
影响因子：
3.3
作者：
Bagchi, Biman;Blainey, Paul C.;Xie, X. Sunney
通讯作者：
Xie, X. Sunney
影响因子：
15
作者：
Kaiser, Wolfgang;Rant, Ulrich
通讯作者：
Rant, Ulrich
影响因子：
38.3
作者：
通讯作者：
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