Unfolding and Translocation of Proteins Through an Alpha-Hemolysin Nanopore by ClpXP

Part of the Methods in Molecular Biology book series (MIMB, volume 2186)


Proteins present a significant challenge for nanopore-based sequence analysis. This is partly due to their stable tertiary structures that must be unfolded for linear translocation, and the absence of regular charge density. To address these challenges, here we describe how ClpXP, an ATP-dependent protein unfoldase, can be harnessed to unfold and processively translocate multi-domain protein substrates through an alpha-hemolysin nanopore sensor. This process results in ionic current patterns that are diagnostic of protein sequence and structure at the single-molecule level.

Key words

Nanopore Protein folding Protein sequencing Single-molecule Unfoldase α-Hemolysin ClpXP 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  1. 1.Paul G. Allen School of Computer Science and EngineeringUniversity of WashingtonSeattleUSA
  2. 2.UC Santa Cruz Genomics InstituteUniversity of California, Santa CruzSanta CruzUSA
  3. 3.Department of Chemistry and BiochemistryUniversity of Notre DameNotre DameUSA

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