Expression and purification of soluble human APRIL in Escherichia coli using ELP-SUMO tag

被引：11

作者：

Zhang, Jie ^{[1
]}

Ma, Lei ^{[2
]}

Zhang, Shuang Quan ^{[2
]}

机构：

[1] Soochow Univ, Affiliated Hosp 1, Jiangsu Inst Hematol, Key Lab Thrombosis & Hemostasis,Minist Hlth, Suzhou, Jiangsu, Peoples R China

[2] Nanjing Normal Univ, Life Sci Coll, Jiangsu Prov Key Lab Mol & Med Biotechnol, Nanjing 210046, Jiangsu, Peoples R China

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2014年 / 95卷

关键词：

ELP-SUMO; hAPRIL; Escherichia coli; Soluble expression; PROTEIN-PURIFICATION; FUSION; SYSTEM; CELL; RECEPTOR; FAMILY; TACI; BCMA;

D O I：

10.1016/j.pep.2013.12.013

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

APRIL is a member of the tumor necrosis factor (TNF) family of ligands that mediate tumor cells proliferation as well as survival, depending on the cellular context. In this report, we present a novel method to obtain soluble human APRIL in Escherichia coli using the elastin-like polypeptide and SUMO (ELP-SUMO) tags. The fusion protein with ELP-SUMO tag was expressed in a soluble form at 15 degrees C. After purification based on inverse transition cycling (ITC) method, the purified ELP-SUMO-hAPRIL fusion protein was subsequently cleaved by SUMO protease to release mature hAPRIL. Following affinity chromatography, the target protein was re-purified with high purity. Finally, about 4.8 mg recombinant hAPRIL was obtained from 11 bacterial culture with no less than 85% purity. The molecular mass (Mr) of the recombinant hAPRIL was confirmed by MALDI-TOF MS as Mr 16,314. The purified hAPRIL exhibits biological activity on Jurkat cells. It is the first report on soluble production of hAPRIL in E. coli using ELP-SUMO tag. (C) 2014 Elsevier Inc. All rights reserved.

引用

页码：177 / 181

页数：5

共 20 条

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