利用燕麦氨肽酶1（OaAEP1）和烟草蚀纹病毒蛋白酶（TEV蛋白酶）进行蛋白质聚合物/多蛋白的酶促构建。
Enzymatic Construction of Protein Polymer/Polyprotein Using OaAEP1 and TEV Protease.
作者信息
Deng Yibing, Shi Shengchao, Zheng Bin, Wu Tao, Zheng Peng
机构信息
State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu, 21002, China.
出版信息
Bio Protoc. 2020 Apr 20;10(8):e3596. doi: 10.21769/BioProtoc.3596.
The development of chemical and biological coupling technologies in recent years has made possible of protein polymers engineering. We have developed an enzymatic method for building polyproteins using a protein ligase OaAEP1 (asparagine endopeptidase 1) and protease TEV (tobacco etching virus). Using a mobile TEV protease site compatible with the OaAEP1 ligation, we achieved a stepwise polymerization of the protein on the surface. The produced polyprotein can be verified by protein unfolding scenario using atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS). Thus, this study provides an alternative method for polyprotein engineering and immobilization.
近年来，化学和生物偶联技术的发展使蛋白质聚合物工程成为可能。我们开发了一种酶促方法，利用蛋白质连接酶OaAEP1（天冬酰胺内肽酶1）和蛋白酶TEV（烟草蚀纹病毒）构建多聚蛋白。通过使用与OaAEP1连接兼容的可移动TEV蛋白酶位点，我们在表面实现了蛋白质的逐步聚合。所产生的多聚蛋白可以通过基于原子力显微镜的单分子力谱（AFM-SMFS）使用蛋白质解折叠方案进行验证。因此，本研究为多聚蛋白工程和固定化提供了一种替代方法。
Zotero 插件