Nanopore-based protein sequencing


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You have full access to this article via your institution. Download PDF Nivala, J. _et al_. _Nat. Biotechnol._ 31, 247–250 (2013). Nanopore-based DNA sequencing is well on its way to


becoming a practical technique. Such an approach could also be adapted to sequence proteins. Before a bench-top protein sequencer can be made, however, several methodological challenges must


be overcome. In new work, Nivala _et al_. address two of these challenges: how to unfold the protein to allow it to go through the pore, and how to ensure that it goes through the pore


processively and unidirectionally. The researchers created a system using the α-hemolysin pore with the unfoldase ClpX positioned on the _trans_ side of the pore. They designed a polyanionic


tag (for the protein of interest) that is captured and threaded through the pore when a voltage is applied; ClpX then recognizes a targeting motif on the C terminus of the tag and, fueled


by ATP hydrolysis, unfolds and pulls the protein of interest through the pore. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Nanopore-based protein


sequencing. _Nat Methods_ 10, 285 (2013). https://doi.org/10.1038/nmeth.2417 Download citation * Published: 28 March 2013 * Issue Date: April 2013 * DOI: https://doi.org/10.1038/nmeth.2417


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