Protein sensing with engineered protein nanopores.
Protein sensing with engineered protein nanopores.
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DOI：
10.1007/978-1-61779-773-6_2
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发表时间：
2012-01-01
期刊：
影响因子：
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通讯作者：
Movileanu, Liviu
中科院分区：
文献类型：
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作者：
Mohammad, Mohammad M;Movileanu, Liviu
纳米孔的使用是单分子科学的一个强大的新前沿。纳米孔已被有效地用于探索小多肽和蛋白质的各种生物物理特征，例如它们的折叠状态和结构、配体相互作用和酶活性。特别地，α-溶血素（alphaHL）蛋白质孔已被广泛用于多肽的检测、表征和分析，因为这种蛋白质纳米孔在各种实验条件下是高度稳健的、通用的和易处理的。受线粒体外膜易位酶中蛋白质易位机制的启发，我们已经证明了使用纳米孔探针技术控制单个蛋白质的能力。在这里，我们提供了一个详细的方案，用于制备α HL蛋白纳米孔。此外，我们表明，放置有吸引力的静电陷阱是有助于解决折叠蛋白质的单分子随机传感。
The use of nanopores is a powerful new frontier in single-molecule sciences. Nanopores have been used effectively in exploring various biophysical features of small polypeptides and proteins, such as their folding state and structure, ligand interactions, and enzymatic activity. In particular, the alpha-hemolysin (alphaHL) protein pore has been used extensively for the detection, characterization, and analysis of polypeptides because this protein nanopore is highly robust, versatile, and tractable under various experimental conditions. Inspired by the mechanisms of protein translocation across the outer membrane translocases of mitochondria, we have shown the ability to use nanopore-probe techniques in controlling a single protein using engineered alphaHL pores. Here, we provide a detailed protocol for the preparation of alphaHL protein nanopores. Moreover, we demonstrate that placing attractive electrostatic traps is instrumental in tackling single-molecule stochastic sensing of folded proteins.