Abstract

Foremost among the challenges facing single molecule sequencing of proteins by nanopores is the lack of a universal method for driving proteins or peptides into nanopores. In contrast to nucleic acids, the backbones of which are uniformly negatively charged nucleotides, proteins carry positive, negative and neutral side chains that are randomly distributed. Recombinant proteins carrying a negatively charged oligonucleotide or polypeptide at the C-termini can be translocated through a α-hemolysin (α-HL) nanopore, but the required genetic engineering limits the generality of these approaches. In this present study, we have developed a chemical approach for addition of a charged oligomer to peptides so that they can be translocated through nanopores. As an example, an oligonucleotide PolyT20 was tethered to peptides through first selectively functionalizing their N-termini with azide followed by a click reaction. The data show that the peptide-PolyT20 conjugates translocated through nanopores, whereas the unmodified peptides did not. Surprisingly, the conjugates with their peptides tethered at the 5′-end of PolyT20 passed the nanopores more rapidly than the PolyT20 alone. The PolyT20 also yielded a wider distribution of blockade currents. The same broad distribution was found for a conjugate with its peptide tethered at the 3′-end of PolyT20, suggesting that the larger blockades (and longer translocation times) are associated with events in which the 5′-end of the PolyT20 enters the pore first.

Original languageEnglish (US)
Pages (from-to)9652-9664
Number of pages13
JournalACS Nano
Volume9
Issue number10
DOIs
StatePublished - Sep 12 2015

Fingerprint

Nanopores
threads
Peptides
peptides
DNA
deoxyribonucleic acid
solid state
proteins
oligonucleotides
Oligonucleotides
Proteins
Genetic engineering
Recombinant proteins
sequencing
Polypeptides
Nucleic acids
polypeptides
nucleotides
nucleic acids
Hemolysin Proteins

Keywords

  • click addition
  • DNA thread
  • nanopore
  • peptide translocation
  • peptide-PolyT conjugate
  • protein sequencing

ASJC Scopus subject areas

  • Engineering(all)
  • Materials Science(all)
  • Physics and Astronomy(all)

Cite this

Click Addition of a DNA Thread to the N-Termini of Peptides for Their Translocation through Solid-State Nanopores. / Biswas, Sudipta; Song, Weisi; Borges, Chad; Lindsay, Stuart; Zhang, Peiming.

In: ACS Nano, Vol. 9, No. 10, 12.09.2015, p. 9652-9664.

Research output: Contribution to journalArticle

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