| 大鼠lipocalin 11蛋白非保守半胱氨酸双点突变体样品制备及理化性质表征 |
| |
| 引用本文： | 顾轶娜,林东海.大鼠lipocalin 11蛋白非保守半胱氨酸双点突变体样品制备及理化性质表征[J].药物生物技术,2011(3):189-194. |
| |
| 作者姓名： | 顾轶娜 林东海 |
| |
| 作者单位： | 中国药科大学生命科学与技术学院;厦门大学化学化工学院;中国科学院上海药物研究所; |
| |
| 基金项目： | 国家自然科学基金项目(No.30730026); 上海市优秀学科带头人项目(No.09XD1405100) |
| |
| 摘 要： | Lipocalin 11蛋白是新发现的一个lipocalin分泌型蛋白家族成员,与男性生殖密切相关。大鼠lipocalin 11(rLcn11)蛋白中4个半胱氨酸往往不能正确配对,为重组蛋白样品的制备带来困难。该研究通过序列比对和同源模建的方法获得rLcn11蛋白非保守半胱氨酸的位点,构建了rLcn11蛋白野生型与C59A/C156A突变体的表达系统和纯化方法。通过分子筛和圆二色谱分析rLcn11蛋白的聚集形态和结构特征,并用荧光滴定的方法研究了rLcn11蛋白与荧光探针ANS的结合能力。结果表明:(1)rLcn11蛋白非保守半胱氨酸残基的突变有利于减少蛋白的错误折叠和聚集,提高重组蛋白的产量;(2)rLcn11 C59A/C156A突变体的空间结构良好并能结合疏水性配体;(3)rLcn11蛋白Cys59、Cys156双点突变不显著地改变rLcn11蛋白的空间结构及其与配体的结合能力。该工作为rLcn11蛋白的空间结构解析和生理功能的研究奠定了基础。
|
| 关 键 词： | Lipocalin11蛋白 半胱氨酸突变 样品制备 配体结合 |
Sample Preparation and Physicochemical Characterization of Rat Lipocalin 11 Nonconservative Cysteins Double Mutant |
| |
| GU Yi-na,LIN Dong-hai,.Sample Preparation and Physicochemical Characterization of Rat Lipocalin 11 Nonconservative Cysteins Double Mutant[J].Pharmaceutical Biotechnology,2011(3):189-194. |
| |
| Authors: | GU Yi-na LIN Dong-hai |
| |
| Institution: | GU Yi-na1,LIN Dong-hai1,2,3(1.School of Life Science and Technology,China Pharmaceutical University,Nanjing 210009,China,2.The Key Laboratory for Chemical Biology of Fujian Province,Department of Chemical Biology,School of Chemistry and Chemical Engineering,Xiamen University,Xiamen 361005,3.Shanghai Institute of Materia Medica,Chinese Academy of Science,Shanghai 201203,China) |
| |
| Abstract: | Lipocalin 11 is a novel secretory protein of the lipocalin family and is potentially related to male reproduction.Misfolding due to the four wrongly paired cysteins in rat lipocalin 11(rLcn11) makes it difficult to prepare the recombinant protein sample.The present work in our lab is to explore two nonconservative cysteins in rLcn11 using sequence alignment and homology modeling.The plasmids of the wild type rLcn11 and its C59A/C156A mutant were constructed and then expressed and purified rLcn11 proteins in... |
| |
| Keywords: | Lipocalin 11 Cysteins mutations Sample preparation Ligand-binding |
| 本文献已被 CNKI 等数据库收录！ |
