Detection of benzo[a]pyrene-guanine adducts in single-stranded DNA using the α-hemolysin nanopore

Perera, Rukshan T. and Fleming, Aaron M. and Johnson, Robert P. and Burrows, Cynthia J. and White, Henry S. (2015) Detection of benzo[a]pyrene-guanine adducts in single-stranded DNA using the α-hemolysin nanopore. Nanotechnology, 26 (7). 074002. ISSN 0957-4484

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Item Type:Article
Item Status:Live Archive

Abstract

The carcinogenic precursor benzo[a]pyrene (BP), a polycyclic aromatic hydrocarbon, is released into the environment through the incomplete combustion of hydrocarbons. Metabolism of BP in the human body yields a potent alkylating agent (benzo[a]pyrene diol epoxide, BPDE) that reacts with guanine (G) in DNA to form an adduct implicated in cancer initiation. We report that the α-hemolysin (αHL) nanopore platform can be used to detect a BPDE adduct to G in synthetic oligodeoxynucleotides. Translocation of a 41-mer poly-2'-deoxycytidine strand with a centrally located BPDE adduct to G through αHL in 1 M KCl produces a unique multi-level current signature allowing the adduct to be detected. This readily distinguishable current modulation was observed when the BPDE-adducted DNA strand translocated from either the 5' or 3' directions. This study suggests that BPDE adducts and other large aromatic biomarkers can be detected with αHL, presenting opportunities for the monitoring, quantification, and sequencing of mutagenic compounds from cellular DNA samples.

Keywords:benzo[a]pyrene, nanopore, hemolysin, NotOAChecked
Subjects:F Physical Sciences > F170 Physical Chemistry
Divisions:College of Science > School of Chemistry
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ID Code:27424
Deposited On:03 May 2017 11:15

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