期刊
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
卷 181, 期 3, 页码 939-947出版社
HUMANA PRESS INC
DOI: 10.1007/s12010-016-2260-z
关键词
Tagremoval; Stericocclusions; Maltosebinding protein; Integral membrane protein; Expression and purification; TEV protease
资金
- Ministry of Science and Technology [2016YFA0501200]
- National Natural Science Foundation of China [31500603, 21425523]
- Fundamental Research Funds for the Central Universities [WUT 2016IB006]
Tag removal is a prerequisite issue for structural and functional analysis of affinity-purified membrane proteins. The present study took a MBP-fused membrane protein, MrpF, as a model to investigate the tag removal by TEV protease. Influences of the linking sequence between TEV cleavage site and MrpF on protein expression and predicted secondary structure were investigated. The steric accessibility of TEV protease to cleavage site of MBP-fused MrpF was explored. It was found that reducing the size of hydrophilic group of detergents and/or extending the linking sequence between cleavage site and target protein can significantly improve the accessibility of the cleavage site and promote tag removal by TEV protease.
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