Research | Polypeptide translocation and sequencing

We have engineered many variants of the α-hemolysin protein pore for single molecule detection (stochastic sensing), which is effected by monitoring the modulation of an ionic current passing through a single pore. Stochastic sensing allows the analysis of a wide variety of analytes: metal cations, small organic molecules, nucleic acids, proteins, and chemicals. The spin-out company from the Bayley laboratory, Oxford Nanopore, has used this technology to sequence single molecules of DNA and RNA. Following the success of Oxford Nanopore's MinION sequencer, we have redirected our efforts towards protein characterization, in particular the detection of post-translational modifications and alternative splicing. To pass through a nanopore, a protein must unfold so that the polypeptide chain can thread through the narrow lumen. Accordingly, this process is also revealing fundamental information about protein folding and the trafficking of proteins between compartments in cells. A critical issue in stochastic sensing is the implementation of highly parallel analyte detection, and we are currently developing technologies to allow simultaneous recording from thousands of protein nanopores.

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Research papers:

Rosen, C.B., Rodriguez-Larrea, D. and Bayley, H., Single-molecule site-specific detection of protein phosphorylation. Nature Biotechnology, 32, 179 (2014). DOI:10.1038/nnano.2013.22

Huang, S., Romero-Ruiz, M., Castell, O.K., Bayley, H., and Wallace, M.I., High-throughput optical sensing of nucleic acids in a nanopore array. Nature Nanotechnology, 10, 986, (2015). DOI:10.1038/nnano.2015.189

Review:

Bayley, H., Nanopore sequencing: from imagination to reality., Clinical Chemistry, 61, 25 (2015). DOI:10.1373/clinchem.2014.223016