Molecular bases of cyclodextrin adapter interactions with engineered protein nanopores.
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 (alphaHL) pore that bind the adapter beta-cyclodextrin (betaCD) approximately 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.
Banerjee
Arijit
A
Department of Chemistry, University of Oxford, Oxford, OX1 3TA, United Kingdom.
Mikhailova
Ellina
E
Cheley
Stephen
S
Gu
Li-Qun
LQ
Montoya
Michelle
M
Nagaoka
Yasuo
Y
Gouaux
Eric
E
Bayley
Hagan
H
D018360:Crystallography, X-Ray; D003505:Cyclodextrins / Q000737:chemistry*; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000235:genetics; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D008958:Models, Molecular; D009154:Mutation; D049329:Nanostructures / Q000737:chemistry*; D016062:Porosity; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013816:Thermodynamics
D003505:Cyclodextrins; D006460:Hemolysin Proteins; D011994:Recombinant Proteins
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