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学科主题: 基础医学
题名:
Soluble expression in Escherichia coli, purification and characterization of a human TF-1 cell apoptosis-related protein TFAR19
作者: Feng, YM; Zhang, YM; Jing, GZ
刊名: PROTEIN EXPRESSION AND PURIFICATION
发表日期: 2002-07-01
卷: 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
Citation statistics:
内容类型: 期刊论文
版本: 出版稿
URI标识: http://ir.bjmu.edu.cn/handle/400002259/64729
Appears in Collections:基础医学院_免疫学系_期刊论文

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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

Recommended Citation:
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.
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