title: Investigation of MTH1 activity via mismatch-based DNA chain elongation.
abstract: Accumulation and misincorporation of oxidative damaged 8-oxo-7,8-dihydroguanine triphosphates (8-oxo-dGTP) in genomic DNA may cause serious cellular function disorders. MutT Homolog 1 (MTH1), a protein enzyme that can help to prevent 8-oxo-dGTP misincorporation, plays critical roles in oxidative stress neutralization, oncogene-associated tumor malignancy, and anticancer therapies. So, in this work, a simple and function-oriented method is developed for the assay of MTH1 activity. Specifically, a mismatch-based ("8-oxoG: A" mismatch) DNA chain elongation strategy (MB-DCE) is firstly proposed to reveal the misincorporation efficiency of 8-oxo-dGTP. Then, further coupled with the inherent activity of MTH1 to prevent 8-oxo-dGTP misincorporation, a relationship can be established to reveal the activity of MTH1 through MB-DCE. As the method is designed directly towards the cellular function of MTH1, activity of MTH1 in different breast cancer cell lines has been detected, implying the potential application of this assay method for biomedical research and clinical diagnose in the future.
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, Collaborative Innovation Center of Chemistry for Life Sciences, Department of Biochemistry, Nanjing University, Nanjing 210093, PR China.
authors.forename: Tao
authors.initials: T
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, Collaborative Innovation Center of Chemistry for Life Sciences, Department of Biochemistry, Nanjing University, Nanjing 210093, PR China.
authors.forename: Shiyu
authors.initials: S
authors.lastname: Gu
authors.affiliation: Department of Oncology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, PR China.
authors.forename: Fengzhen
authors.initials: F
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, Collaborative Innovation Center of Chemistry for Life Sciences, Department of Biochemistry, Nanjing University, Nanjing 210093, PR China.
authors.forename: Liudi
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Oncology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, PR China.
authors.forename: Zhaoxia
authors.initials: Z
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, Collaborative Innovation Center of Chemistry for Life Sciences, Department of Biochemistry, Nanjing University, Nanjing 210093, PR China.
authors.forename: Jie
authors.initials: J
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, Collaborative Innovation Center of Chemistry for Life Sciences, Department of Biochemistry, Nanjing University, Nanjing 210093, PR China; Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, PR China. Electronic address: genxili@nju.edu.cn.
authors.forename: Genxi
authors.initials: G
authors.lastname: Li
mesh_terms: D020137:Base Pair Mismatch; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry*; D045643:DNA Repair Enzymes / Q000378:metabolism*; D004566:Electrodes; D006801:Humans; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*
chemical_list: D004247:DNA; D010744:Phosphoric Monoester Hydrolases; C078207:8-oxodGTPase; D045643:DNA Repair Enzymes
keywords: DNA chain elongation; Electrochemical assay; MutT Homolog 1 activity; Nucleobase misincorporation; Oxidative damages