Thursday, February 14, 2013
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Assays & screens
through an a-hemolysin-caseinolytic peptidase X homolog (ClpX)
A method to translocate
proteins through a-hemolysin nanopores could eventually enable
nanopore-based protein sequencing. There are nanopore-based DNA sequencing
methods in commercial development that detect and identify DNA by measuring
voltage changes as individual base pairs pass through a membrane-embedded
nanopore, but these methods cannot identify peptides. To enable protein
translocation through a membrane-embedded nanopore, the AAA+ unfoldase ClpX was added in solution to one side of
the membrane, allowing detection of structure-dependent voltage changes as a
protein substrate of about 100 amino acids and carrying a ClpX binding tag
passed through the pore. Next steps could include determining the relationship
between voltage change and amino acid identity.
At least five companies are developing nanopore-based DNA sequencing systems.
Published online Feb. 14, 2013
Patent and licensing status
Nivala, J. et al. Nat.
Biotechnol.; published online Feb. 3, 2013;
Contact: Mark Akeson, University of California, Santa Cruz, Calif.
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