文献类型：期刊文献

英文题名：Highly Efficient Cofactors of Cu2+-Dependent Deoxyribozymes

作者：Han, Haiyan[1];Huang, Qiping[1];Liu, Hui[1];Zhang, Jingyan[1,2,3]

机构：[1]East China Univ Sci & Technol, Sch Pharm, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Univ Minnesota, Dept Biochem Mol Biol & Biophys, Minneapolis, MN 55455 USA;[3]Univ Minnesota, Ctr Met Biocatalysis, Minneapolis, MN 55455 USA

年份：2017

卷号：2

期号：13

起止页码：3925

外文期刊名：CHEMISTRYSELECT

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000401890300047)】；

基金：We thank Prof. John D. Lipscomb (University of Minnesota) for his comments and advice on the manuscript. This work was supported by the state key laboratory of bioreactor engineering [No. 2060204], 111 Project [No. B07023], National Nature Science Foundation [No. 21671065] and the Shanghai Committee of Science and Technology [No. 11DZ2260600].

语种：英文

外文关键词：Cu2+-dependent DNAzyme; Cofactor; Cu2+; Copper compounds; DNA cleavage

摘要：Deoxyribozymes can catalyze various types of reactions mostly with metal ions as their cofactors. Here we demonstrate that three copper compounds with thiosemicarbazide or semicarbazide ligands, are highly active cofactors of Cu2+-dependent deoxyribozymes without any additional reagents. The maximum catalytic rate constant of the deoxyribozyme with dichloro(di-thiosemicarbazide) copper as a cofactor is similar to 25-fold faster than Cu2+ cofactor under the same reaction condition. Using a variety of spectroscopies, electrochemistry, and mutagenesis, we demonstrate that both the three-dimensional structure and redox potential of the copper center of the cofactors are essential to the catalysis of deoxyribozymes. The cofactor interacts with the enzyme-substrate complex forming a ternary enzyme-substrate-cofactor complex, which is confirmed by changing the structure of the enzyme-substrate complex through varying the temperature, or mutating the active site of the enzyme and conserved site of the substrate. The result suggests that Cu2+-dependent deoxyribozyme requires a well-defined active site to carry out the catalysis proficiently.