Detection of benzo[a]pyrene-guanine adducts in single-stranded DNA using the α-hemolysin nanopore.
Detection of benzo[a]pyrene-guanine adducts in single-stranded DNA using the α-hemolysin nanopore.
复制标题
使用α-脱糖蛋白纳米孔检测单链DNA中苯并[A] pyrene-rene-rene加合物。
DOI：
10.1088/0957-4484/26/7/074002
复制
发表时间：
2015-02-20
期刊：
影响因子：
3.5
通讯作者：
White HS
中科院分区：
文献类型：
--
作者：
Perera RT;Fleming AM;Johnson RP;Burrows CJ;White HS
致癌前体苯并[a]芘（BP）是一种多环芳烃，通过碳氢化合物的不完全燃烧释放到环境中。 BP 在人体内的代谢产生一种有效的烷化剂（苯并[a]芘二醇环氧化物，BPDE），它与 DNA 中的鸟嘌呤 (G) 反应形成与癌症发生有关的加合物。我们报告称，α-溶血素 (αHL) 纳米孔平台可用于检测合成寡脱氧核苷酸中 BPDE 与 G 的加合物。 41 聚体聚 2'-脱氧胞苷链与位于中心的 BPDE 加合物通过 1 M KCl 中的 αHL 易位至 G，产生独特的多级电流特征，允许检测加合物。当 BPDE 加合 DNA 链从 5' 或 3' 方向易位时，观察到这种易于区分的电流调制。这项研究表明，BPDE 加合物和其他大型芳香族生物标志物可以用 αHL 进行检测，为细胞 DNA 样本中的诱变化合物的监测、定量和测序提供了机会。
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.