title: Detection of benzo[a]pyrene-guanine adducts in single-stranded DNA using the α-hemolysin nanopore.
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.
authors: Department of Chemistry, University of Utah, 315 South 1400 East, Salt Lake City, Utah 84112-0850, USA.
authors: Rukshan T
authors: RT
authors: Perera
authors: Aaron M
authors: AM
authors: Fleming
authors: Robert P
authors: RP
authors: Johnson
authors: Cynthia J
authors: CJ
authors: Burrows
authors: Henry S
authors: HS
authors: White
mesh_terms: D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000737:chemistry*; D000477:Alkylating Agents / Q000737:chemistry; D001564:Benzo(a)pyrene / Q000737:chemistry*; D018736:DNA Adducts / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry*; D016678:Genome; D005898:Glass; D006147:Guanine / Q000737:chemistry*; D006460:Hemolysin Proteins / Q000737:chemistry*; D006801:Humans; D007477:Ions / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D008670:Metals / Q000737:chemistry; D016296:Mutagenesis; D009153:Mutagens; D058608:Nanopores; D036103:Nanotechnology; D009369:Neoplasms / Q000378:metabolism*; D009841:Oligonucleotides / Q000737:chemistry
chemical_list: D000477:Alkylating Agents; D018736:DNA Adducts; D004277:DNA, Single-Stranded; D006460:Hemolysin Proteins; D007477:Ions; D008051:Lipid Bilayers; D008670:Metals; D009153:Mutagens; D009841:Oligonucleotides; C043931:benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; D001564:Benzo(a)pyrene; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D006147:Guanine
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