title: DNA methyltransferase detection based on digestion triggering the combination of poly adenine DNA with gold nanoparticles.
abstract: DNA methyltransferase (MTase) has received a large amount of attention due to its catalyzation of DNA methylation in both eukaryotes and prokaryotes, which has a close relationship to cancer and bacterial diseases. Herein, a novel electrochemical strategy based on Dpn I digestion triggering the combination of poly adenine (polyA) DNA with a gold nanoparticles functioned glassy carbon electrode (AuNPs/GCE), is developed for the simple and efficient detection of DNA MTase and inhibitor screening. Only one methylene blue (MB)-labeled DNA hairpin probe and two enzymes are involved in this designed method. In the presence of Dam MTase, the hairpin probe can be methylated and then cleaved by the restriction endonuclease. Thus, a MB-labeled polyA signal-stranded DNA product is introduced to the surface of AuNPs/GCE through the effect between polyA and AuNPs, resulting in an obvious electrochemical signal. On the contrary, in the absence of Dam MTase, the DNA probe cannot be cleaved and a relatively small electrochemical response can be observed. As a result, the as-proposed biosensor offered an efficient way for Dam MTase activity monitoring with a low detection of 0.27 U/mL, a wide linear range and good stability. Additionally, this assay holds great potential for further application in real biological matrices and inhibitors screening, which is expected to be useful in disease diagnosis and drug discovery.
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, PR China.
authors.forename: Pei
authors.initials: P
authors.lastname: Liu
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, PR China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Wang
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, PR China.
authors.forename: Yunlei
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, PR China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Wang
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, PR China. Electronic address: yinhs@sdau.edu.cn.
authors.forename: Huanshun
authors.initials: H
authors.lastname: Yin
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, PR China. Electronic address: ashy@sdau.edu.cn.
authors.forename: Shiyun
authors.initials: S
authors.lastname: Ai
mesh_terms: D000225:Adenine / Q000737:chemistry*; D015374:Biosensing Techniques; D019175:DNA Methylation / Q000235:genetics*; D015254:DNA Modification Methylases / Q000737:chemistry / Q000302:isolation & purification*; D006046:Gold / Q000737:chemistry; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry
chemical_list: D006046:Gold; D015254:DNA Modification Methylases; D000225:Adenine
keywords: DNA methyltransferase; Electrochemical biosensor; Inhibitor; Poly adenine