| 通过快速定点突变表征人源含硒单链抗体酶的底物结合部位 |
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| 引用本文： | 宋健,徐俊杰,魏景艳,于扬,孙卫国,张桂荣.通过快速定点突变表征人源含硒单链抗体酶的底物结合部位[J].高等学校化学学报,2010,31(3):502. |
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| 作者姓名： | 宋健 徐俊杰 魏景艳 于扬 孙卫国 张桂荣 |
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| 作者单位： | 1. 吉林大学电子科学与工程学院,
2. 白求恩医学院,
3. 药学院, 长春 130021;
4. 吉林医药学院基础医学院, 吉林 132013 |
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| 基金项目： | 国家自然科学基金(批准号:30870540);;吉林省科技发展计划项目(批准号:20070726,20070425,200705368)资助 |
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| 摘 要： | 为了对人源含硒单链抗体酶Se-scFv-B3的底物结合部位和催化基团进行研究, 在理论预测的基础上, 通过快速定点突变法分别在2个理论预测的底物结合部位(位点1和位点2)内选定Ala180和Ala44定点突变为丝氨酸(Ser). 2个突变体蛋白经化学修饰将Ser转变成谷胱甘肽过氧化物酶(GPX)的催化基团硒代半胱氨酸(Sec)后, 前者的GPX活力达到了Se-scFv-B3的2倍多, 而后者的GPX活力没有明显提高, 这表明位点1可能是主要的底物结合部位, 与理论预测的结果一致.
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| 关 键 词： | 快速定点突变 人源含硒单链抗体酶 底物结合部位 |
| 收稿时间： | 2009-05-14 |
Substrate-binding Sites of Human Selenium-containing Single Chain Abzyme by Quikchange Site-directed Mutagenesis |
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| SONG Jian,XU Jun-Jie,WEI Jing-Yan,YU Yang,SUN Wei-Guo,ZHANG Gui-Rong.Substrate-binding Sites of Human Selenium-containing Single Chain Abzyme by Quikchange Site-directed Mutagenesis[J].Chemical Research In Chinese Universities,2010,31(3):502. |
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| Authors: | SONG Jian XU Jun-Jie WEI Jing-Yan YU Yang SUN Wei-Guo ZHANG Gui-Rong |
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| Institution: | 1. College of Electronic Science and Engineering, ;
2. College of Fundamental Medicine,;
3. College of Pharmaceutical Science; Jilin University, Changchun 130021, China;
4. Department of Biochemistry, School of Basic Medical Sciences, Jilin Medicine College, Jilin 132013, China |
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| Abstract: | Ala180 in site 1 and Ala44 in site 2 of human selenium-containing single chain abzyme Se-scFv-B3 were chosen to be mutated to serines,respectively,to study the substrate binding sites and the catalytic group of Se-scFv-B3 and to find the reason why the Se-scFv-B3 has lower catalytic activity based on the theoretical anticipation for the protein.After serine residue was reacted into secystins by chemical modification,the activity of the anterior one is about two times than that of the Se-scFv-B3,but the latt... |
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| Keywords: | Quikchange site-directed mutagenesis Human selenium-containing sigle chain abzyme Substrate-binding site |
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