A versatile platform for single-molecule enzymology of restriction endonuclease
作者机构：National Laboratory of Solid State Microstructure School of PhysicsNanjing University 22 Hankou Road NanjingJiangsu 210093P.R.China School of Chemistry and Chemical Engineering Nanjing University 163 Xianlin Avenue NanjingJiangsu 210023P.R.China
出 版 物：《Journal of Innovative Optical Health Sciences》 (创新光学健康科学杂志（英文）)
年 卷 期：2019年第12卷第1期
页 面：29-38页
核心收录：
学科分类：0831[工学-生物医学工程（可授工学、理学、医学学位）] 08[工学] 0836[工学-生物工程]
基 金：The authors greatly appreciate the-nancial support from National Natural Science Foundation of China(Grant Nos.21522402,11674153,11374148,11334004 and 21771103) the Fundamental Research Funds for the Central Universities(Nos.020414380070,020414380050 and 020414380058) Natural Science Foundation of Jiangsu Province(No.BK20160639)and the Shuangchuang Program of Jiangsu Province
主 题：Single molecule Forster resonance energy transfer enzymology kinetics holliday junction
摘 要：Enzymes are the major players for many biological *** studies of the enzymatic activity at the single-molecule level provides important information that is otherwise inaccessible at the ensemble ***,these single-molecule experiments are technically di±cult and generally require complicated experimental ***,we develop a Holliday junction(HJ)-based platform to study the activity of restriction endonucleases at the single-molecule level using single-molecule FRET(sm-FRET).We show that the intrinsic dynamics of HJ can be used as the reporter for both the enzyme-binding and the substrate-release *** to the multiple-arms structure of HJ,the fluorophore-labeled arms can be different from the surface anchoring arm and the substrate ***,it is possible to independently change the substrate arm to study different enzymes with similar *** a design is extremely useful for the systematic study of enzymes from the same family or enzymes bearing different pathologic ***,this method can be easily extended to study other types of DNA-binding enzymes without too much modi fication of the *** anticipate it can find broad applications in single-molecule enzymology.
维普期刊数据库