Novel Selenium-containing Human Single-chain Variable Fragment with Glutathione Peroxidase Activity from Computer-aided Molecular Design

作     者：WANG Cheng WAN Pei GONG Ping-sheng LV Li-min XU Ya-wei ZHAO Yang HE Bo ZHAO Gang YAN Gang-lin MU Ying LV Shao-wu LUO Gui-min 

作者机构：Key Laboratory for Molecular Enzymology and Engineering Ministry of Education College of Life Science Jilin University Changchun 130012 P. R. China State Key Laboratory of Industrial Control Technology Research Center for Analytical Instrumentation Institute of Cyber-systems and Control Zhejiang University Hangzhou 310058 P. R. China Department of Biochemistry School of Basic Medical Sciences Jilin Medical College Jilin 132013 P. R. China 

基　　金：Supported by the National Natural Science Foundation of China(No.30970608) the Applicative Technological Project of Bureau of Science and Technology of Changchun City, China(No.2009045) the Development and Planning Major Program of Jilin Provincial Science and Technology Department, China(No.20100948) the Innovation Method Fund of China (No.2008IM040800) 

出 版 物：《Chemical Research in Chinese Universities》 (高等学校化学研究（英文版）)

年 卷 期：2011年第27卷第5期

页      码：813-819页

摘      要：In order to enhance the glutathione peroxidase（GPX） catalytic activity of the selenium-containing single-chain variable fragments（Se-scFv）, a novel human scFv was designed on the basis of the structure of human antibody and optimized via bioinformatics methods such as homologous sequence analysis, three-dimensional（3D） model building, binding-site analysis and docking. The DNA sequence of the new human scFv was synthesized and cloned into the expression vector pET22b（＋）, then the scFv protein was expressed in soluble form in Escherichia coli BL21（DE3） and purified by Ni2＋-immobilized metal affinity chromatography（IMAC）. The serine residue of scFv in the active site was converted into selenocysteine（Sec） with the chemical modification method, thus, the human Se-scFv with GPX activity was obtained. The GPX activity of the Se-scFv protein was characterized. Compared with other Se-scFv, the new human Se-scFv showed similar efficiency for catalyzing the reduction of hydrogen peroxide by glutathione. It exhibited pH and temperature dependent catalytic activity and a typical ping-pong kinetic mechanism.

主 题 词：Glutathione peroxidase(GPX) Single-chain variable fragment(scFV) Three-dimensional model Selenium 

学科分类：0710[理学-生物科学类] 090502[090502] 071010[071010] 081704[081704] 07[理学] 08[工学] 0817[工学-轻工类] 0905[农学-林学类] 09[农学] 0703[理学-化学类] 

核心收录：

馆 藏 号：203140155...