Molecular bases of cyclodextrin adapter interactions with engineered protein nanopores
Molecular bases of cyclodextrin adapter interactions with engineered protein nanopores
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DOI：
10.1073/pnas.0914229107
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发表时间：
2010-05-04
影响因子：
11.1
通讯作者：
Bayley, Hagan
中科院分区：
文献类型：
--
作者：
Banerjee, Arijit;Mikhailova, Ellina;Bayley, Hagan
工程蛋白质孔在生物技术中有几个潜在的应用：作为随机检测和超快速DNA测序中的传感器元件，作为观察单分子化学的纳米反应器，以及在纳米和微型设备的构建中。一类重要的孔包含分子适配器，其为小分子提供内部结合位点。已经获得了α-溶血素（α HL）孔的突变体，其与接头β-环糊精（β CD）的结合比野生型紧密10（4）倍。我们现在使用单通道电记录，包括非天然氨基酸诱变在内的蛋白质工程和高分辨率X射线晶体学，以无与伦比的细节提供这些工程蛋白质纳米孔的明确结构信息。
Engineered protein pores have several potential applications in biotechnology: as sensor elements in stochastic detection and ultrarapid DNA sequencing, as nanoreactors to observe single-molecule chemistry, and in the construction of nano- and micro-devices. One important class of pores contains molecular adapters, which provide internal binding sites for small molecules. Mutants of the alpha-hemolysin (alpha HL) pore that bind the adapter beta-cyclodextrin (beta CD) similar to 10(4) times more tightly than the wild type have been obtained. We now use single-channel electrical recording, protein engineering including unnatural amino acid mutagenesis, and high-resolution x-ray crystallography to provide definitive structural information on these engineered protein nanopores in unparalleled detail.