胰蛋白酶定向固定化亲和配基的理性设计

引用本文: 白姝, 周荣, 刘夫锋. 胰蛋白酶定向固定化亲和配基的理性设计[J]. 物理化学学报, 2013, 29(02): 439-448. doi: 10.3866/PKU.WHXB201211272 shu

Citation: BAI Shu, ZHOU Rong, LIU Fu-Feng. Rational Design of Affinity Ligand for the Oriented Immobilization of Trypsin[J]. Acta Physico-Chimica Sinica, 2013, 29(02): 439-448. doi: 10.3866/PKU.WHXB201211272 shu

胰蛋白酶定向固定化亲和配基的理性设计

基金项目:
国家自然科学基金(20906068) (20906068)

天津市应用基础与前沿技术研究计划项目(10JCYBJC04500)资助 (10JCYBJC04500)

摘要:

在胰蛋白酶三维(3D)结构的基础上, 首先利用分子对接从ZINC 数据库中筛选获得了与胰蛋白酶具有较高亲和性的小分子配基2-硝基苯基-β-D-葡糖苷, 并分析了该配基与蛋白质之间的相互作用力主要为范德华和氢键相互作用. 并利用分子动力学模拟进一步验证了2-硝基苯基-β-D-葡糖苷与胰蛋白酶之间具有较强的亲和作用. 分子动力学(MD)模拟结果表明, 配基-目标蛋白质之间形成稳定的复合物且它们之间的距离基本没有变化. 此外, 一个水分子通过氢键在配基和目标蛋白质的结合腔之间架桥. 最后制备了偶联有该配基的亲和载体, 进行了胰蛋白酶的定向固定化, 并考察了该固定化酶的活性. 研究结果表明, 利用修饰2-硝基苯基-β-D-葡糖苷配基的亲和载体固定化胰蛋白酶的酶活达到340.8 U·g^-1, 比活达到300.3 U·mg^-1, 分别是未修饰亲和配基载体的10倍和5倍, 具有明显的优势. 上述结论证明了结合分子对接和分子动力学模拟理性设计定向固定化亲和配基的方法是可行的, 具有一定的理论和实用价值.

关键词:

English

Rational Design of Affinity Ligand for the Oriented Immobilization of Trypsin

1.
Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, P. R. China; Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin University, Tianjin 300072, P. R. China
Received Date: 20 September 2012
Available Online: 14 January 2013
Fund Project:
国家自然科学基金(20906068)

天津市应用基础与前沿技术研究计划项目(10JCYBJC04500)资助

Abstract:

Based on the three-dimensional (3D) structure of trypsin, 2-nitrophenyl-β-D-glucopyranoside was selected from the ZINC database to be a candidate affinity ligand for trypsin. The affinity between trypsin and the ligand was analyzed. It is found that the interactions between the ligand and the protein are dominated by van der Waals interactions and hydrogen bonding. Molecular dynamics (MD) simulations were used to verify the affinity between the ligand and trypsin; the simulations indicate that the complex remains stable, and the distance between the ligand and the target protein changes only a little. It is found that one water molecule acts as a bridge between the ligand and the protein pocket via hydrogen bonding. Finally, the ligand was coupled to Sepharose CL-6B gel, and was used to immobilize trypsin in an oriented fashion. It is found that the enzyme activity and specific activity of the oriented immobilized trypsin are 340.8 U·g^-1 and 300.3 U·mg^-1, respectively. These values are 10 and 5 times that of the free enzyme. The results of this work indicate that the combination of docking and MD simulations are promising for the rational design of ligands for oriented immobilization.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

胰蛋白酶定向固定化亲和配基的理性设计

English

Rational Design of Affinity Ligand for the Oriented Immobilization of Trypsin

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

胰蛋白酶定向固定化亲和配基的理性设计

English

Rational Design of Affinity Ligand for the Oriented Immobilization of Trypsin

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs