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 

作者机构：Department of Biomedical EngineeringSchool of EngineeringChina Pharmaceutical UniversityNanjing210009China National Center for Respiratory MedicineCenter of Respiratory MedicineChina-Japan Friendship HospitalBeijing100029China Department of Clinical PharmacyJinling HospitalState Key Laboratory of Analytical Chemistry for Life Science&Jiangsu Key Laboratory of Molecular MedicineMedical School of Nanjing UniversityNanjing210002China School of Pharmaceutical SciencesSouthern Medical UniversityGuangzhou510515China 

出 版 物：《Journal of Analysis and Testing》 (分析检测（英文）)

年 卷 期：2024年第8卷第3期

页      面：327-334页

核心收录：

学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学] 

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) 

主　　题：DNA methyltransferase Methyl-CpG-binding protein Surface plasmon resonance Inhibitor screening Biosensing 

摘      要：DNA methyltransferase(MTase)activity detection has received increasing attention as a promising biomarker and therapeutic ***,most of these detection methods rely on endonuclease digestion and signal groups ***,we present a novel platform for sensing DNA MTase activity that overcomes these *** 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)*** SA-MBD protein specifically recognizes a hairpin probe containing methylated CpG sites,which is treated with *** *** recognition event generates a corresponding SPR response *** limit of detection is as low as 0.016 U/mL,owing to the high-affinity of the SA-MBD ***,we have demonstrated the feasibility of our method for *** MTase activity analysis in serum and inhibitor screening,which implies the potential prospects for biomedical research.