An efficient method for refolding the extracellular portion of CD147   from the total bacterial lysate

doi:10.1093/abbs/gmr060

CORC > 北京大学 > 化学与分子工程学院

	An efficient method for refolding the extracellular portion of CD147 from the total bacterial lysate
	Song, Fei ; Zhang, Xin ; Li, Yifei ; Ru, Qiang ; Ren, Xiaobai ; Xia, Bin ; Chen, Zhi-Nan
刊名	acta biochimica et biophysica sinica
	2011
关键词	CD147 extracellular domain protein refolding isotope labeling nuclear magnetic resonance MATRIX-METALLOPROTEINASE INDUCER T-CELL-ACTIVATION IMMUNOGLOBULIN SUPERFAMILY ESCHERICHIA-COLI RECOMBINANT PROTEINS INCLUSION-BODIES IG SUPERFAMILY MOUSE BASIGIN EXPRESSION EMMPRIN
DOI	10.1093/abbs/gmr060
英文摘要	CD147 is a widely expressed transmembrane protein that mediates signal transduction, and it plays important roles in many physiological and pathological processes, such as tumor invasion and metastasis. The extracellular portion of CD147 (CD147(EC)) is responsible for its functional interactions with different signaling molecules. Due to the existence of two disulfide bonds, CD147(EC) is mainly expressed as an inclusion body in Escherichia coli. Here, we report a convenient rapid-dilution refolding protocol that enables the refolding of CD147(EC) efficiently from total bacterial lysate instead of pure inclusion bodies. Using this method, over 25 mg of CD147(EC) can be purified from 1 l of bacterial culture in M9 medium. The refolded CD147(EC) is well folded as characterized by nuclear magnetic resonance (NMR), and it can induce the expression of matrix metalloproteinase-9 in fibroblast cells. The described protocol is also applicable to the refolding of two immunoglobulin domains of CD147(EC) individually. Interestingly, we noticed that little protein was produced for the C-terminal immunoglobulin (Ig) domain of CD147(EC) by bacteria in M9 medium, even though it was overexpressed in Luria-Bertani (LB) medium. However, when the pH of the bacterial culture in M9 medium was adjusted in accordance with that in LB medium during growth, comparable expression level could be achieved.; Biochemistry & Molecular Biology; Biophysics; SCI(E); 中国科技核心期刊(ISTIC); 中国科学引文数据库(CSCD); 0; ARTICLE; 11; 900-908; 43
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/394338]
专题	化学与分子工程学院生命科学学院
推荐引用方式 GB/T 7714	Song, Fei,Zhang, Xin,Li, Yifei,et al. An efficient method for refolding the extracellular portion of CD147 from the total bacterial lysate[J]. acta biochimica et biophysica sinica,2011.
APA	Song, Fei.,Zhang, Xin.,Li, Yifei.,Ru, Qiang.,Ren, Xiaobai.,...&Chen, Zhi-Nan.(2011).An efficient method for refolding the extracellular portion of CD147 from the total bacterial lysate.acta biochimica et biophysica sinica.
MLA	Song, Fei,et al."An efficient method for refolding the extracellular portion of CD147 from the total bacterial lysate".acta biochimica et biophysica sinica (2011).