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Soluble expression in Escherichia coli, purification and characterization of a human TF-1 cell apoptosis-related protein TFAR19
Feng, YM; Zhang, YM; Jing, GZ
刊名PROTEIN EXPRESSION AND PURIFICATION
2002-07-01
25期:2页:323-329
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
研究领域[WOS]Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
关键词[WOS]DEATH ; BCL-2 ; TRANSITION ; POLYMERASE ; GENES ; BAX
英文摘要

A novel human TF-1 cell apoptosis-related protein, TFAR19, cloned from a human leukemia cell line, TF-1, was first over-expressed in Escherichia coli with the sequence Met-Gly-His(6)-Gly-Thr-Asn-Gly, a hexahistidine sequence followed by a hydroxylamine cleavage site attached to its amino terminus. The resulting protein was soluble and single-step purified to homogeneity by metal chelating affinity chromatography. After cleavage of the purified His(6)-tagged TFAR19 sample with hydroxylamine, highly purified untagged TFAR19 protein was then obtained through an FPLC Resource Q column. The structural characteristics and function of the His(6)-tagged and untagged TFAR19 proteins were studied using circular dichroism, intrinsic fluorescence, and ANS-binding fluorescence spectra and apoptosis activity assay. The results show that alpha-helix is the main secondary structure of the proteins and the two forms of TFAR19 protein fold properly, which correspond well to their apoptosis activity expression. The results also indicate that the extra sequence including the His(6)-tag fused to the N-terminus of TFAR19 protein has a minimal effect on its structure and function, suggesting that the His(6)-tagged TFAR19 protein could be further used as an immobilized target for finding potential proteins which interact with TFAR19 from a cDNA library using in vitro ribosome display technique. (C) 2002 Elsevier Science (USA). All rights reserved.

语种英语
WOS记录号WOS:000177377300016
引用统计
被引频次:6[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
版本出版稿
条目标识符http://ir.bjmu.edu.cn/handle/400002259/64729
专题基础医学院_免疫学系
作者单位1.Acad Sinica, Inst Biophys, Natl Lab Biomacromol, Beijing 100101, Peoples R China
2.Beijing Univ, Hlth Sci Ctr, Lab Med Immunol, Beijing 100083, Peoples R China
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GB/T 7714
Feng, YM,Zhang, YM,Jing, GZ. Soluble expression in Escherichia coli, purification and characterization of a human TF-1 cell apoptosis-related protein TFAR19[J]. PROTEIN EXPRESSION AND PURIFICATION,2002,25(2):323-329.
APA Feng, YM,Zhang, YM,&Jing, GZ.(2002).Soluble expression in Escherichia coli, purification and characterization of a human TF-1 cell apoptosis-related protein TFAR19.PROTEIN EXPRESSION AND PURIFICATION,25(2),323-329.
MLA Feng, YM,et al."Soluble expression in Escherichia coli, purification and characterization of a human TF-1 cell apoptosis-related protein TFAR19".PROTEIN EXPRESSION AND PURIFICATION 25.2(2002):323-329.
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