Enhanced protein expression in the baculovirus/insect cell system using engineered SUMO fusions.
Enhanced protein expression in the baculovirus/insect cell system using engineered SUMO fusions.
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使用工程的Sumo融合，在杆状病毒/昆虫细胞系统中增强了蛋白质的表达。
DOI：
10.1016/j.pep.2008.07.010
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发表时间：
2008-11
影响因子：
1.6
通讯作者：
Strickler, James E.
中科院分区：
文献类型：
--
作者：
Liu, Li;Spurrier, Joshua;Butt, Tauseef R.;Strickler, James E.
Recombinant protein expression in insect cells varies greatly from protein to protein. A fusion tag that is not only a tool for detection and purification, but also enhances expression and/or solubility would greatly facilitate both structure/function studies and therapeutic protein production. We have shown that fusion of SUMO (small ubiquitin-related modifier) to several test proteins leads to enhanced expression levels in E. coli. In eukaryotic expression systems, however, the SUMO tag could be cleaved by endogenous de-SUMOylases. In order to adapt SUMO-fusion technology to these systems, we have developed an alternative SUMO-derived tag, designated SUMOstar, which is not processed by native SUMO proteases. In the present study, we tested the SUMOstar tag in a baculovirus/insect cell system with several proteins, i.e. mouse UBP43, human tryptase beta II, USP4, USP15 and GFP. Our results demonstrate that fusion to SUMOstar enhanced protein expression levels at least 4-fold compared to either the native or His6 tagged proteins. We isolated active SUMOstar tagged UBP43, USP4, USP15, and GFP. Tryptase was active following cleavage with a SUMOstar specific protease. The SUMOstar system will make significant impact in difficult-to-express proteins and especially to those proteins that require the native N-terminal residue for function.
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