A High-Affinity Methyl-CpG-Binding Protein for Endonuclease-Free and Label-Free DNA Methyltransferase Activity Detection  

作　　者：Yang Bai Shulin Tan Yingsong Sheng Yueqing Gu Haiping Wu Baicun Li Yunlong Liu 

机构地区：[1]Department of Biomedical Engineering,School of Engineering,China Pharmaceutical University,Nanjing,210009,China [2]National Center for Respiratory Medicine,Center of Respiratory Medicine,China-Japan Friendship Hospital,Beijing,100029,China [3]Department of Clinical Pharmacy,Jinling Hospital,State Key Laboratory of Analytical Chemistry for Life Science&Jiangsu Key Laboratory of Molecular Medicine,Medical School of Nanjing University,Nanjing,210002,China [4]School of Pharmaceutical Sciences,Southern Medical University,Guangzhou,510515,China

出　　处：《Journal of Analysis and Testing》2024年第3期327-334,共8页分析检测（英文）

基　　金：funded by the National Natural Science Foundation of China(61901527,82300051);National Key Research and Development Program of China(2022YFF0710803,2022YFF0710800);Fundamental Research Funds for the Central Universities(2632021ZD02).

摘　　要：DNA methyltransferase(MTase)activity detection has received increasing attention as a promising biomarker and therapeutic target.However,most of these detection methods rely on endonuclease digestion and signal groups labeling.Herein,we present a novel platform for sensing DNA MTase activity that overcomes these limitations.Our approach is both endonuclease-free and label-free,utilizing a combination of a high-affinity streptavidin-methyl-CpG-binding domain(SA-MBD)protein and surface plasmon resonance(SPR)technology.The SA-MBD protein specifically recognizes a hairpin probe containing methylated CpG sites,which is treated with M.SssI MTase.This recognition event generates a corresponding SPR response signal.The limit of detection is as low as 0.016 U/mL,owing to the high-affinity of the SA-MBD protein.Notably,we have demonstrated the feasibility of our method for M.SssI MTase activity analysis in serum and inhibitor screening,which implies the potential prospects for biomedical research.

关 键 词：DNA methyltransferase Methyl-CpG-binding protein Surface plasmon resonance Inhibitor screening BIOSENSING 

分 类 号：R318[医药卫生—生物医学工程]