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Molecular bases of cyclodextrin adapter interactions with engineered protein nanopores

2010; National Academy of Sciences; Volume: 107; Issue: 18 Linguagem: Inglês

10.1073/pnas.0914229107

1091-6490

Banerjee Ak, Ellina Mikhailova, Stephen Cheley, Li‐Qun Gu, Michelle Montoya, Yasuo Nagaoka, Eric Gouaux, Hagan Bayley,

Microfluidic and Capillary Electrophoresis Applications

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 α-hemolysin (αHL) pore that bind the adapter β-cyclodextrin (βCD) ∼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.