Purification and characterization of recombinant sTRAIL expressed in Escherichia coli.
Purification and characterization of recombinant sTRAIL expressed in Escherichia coli.
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DOI：
10.1093/abbs/36.2.118
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发表时间：
2004-02
影响因子：
3.7
通讯作者：
Xiao-Xia Xia-Xiao-Xia-Xia-4107953;Yaling Shen;D. Wei
中科院分区：
文献类型：
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作者：
Xiao-Xia Xia-Xiao-Xia-Xia-4107953;Yaling Shen;D. Wei
作为一种潜在的抗肿瘤蛋白，肿瘤坏死因子相关凋亡诱导配体（TRAIL）引起了人们的广泛关注。该报告介绍了可溶性 TRAIL 的纯化和表征，在大肠杆菌中以包涵体形式表达。通过简单的稀释方法将sTRAIL包涵体溶解并重折叠至0.9g/L的高浓度。通过一步固定金属亲和层析将重折叠的蛋白质纯化至电泳均质。纯化的sTRAIL对人胰腺肿瘤细胞系1990具有很强的细胞毒活性，ED50约为1.5 mg/L。圆二色性和荧光光谱分析表明，重折叠的sTRAIL具有与天然蛋白相似的β折叠二级结构。这种有效的 sTRAIL 复性程序可能有助于大规模生产这种治疗上重要的蛋白质。
As a potential anti-tumor protein, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has drawn considerable attention. This report presented the purification and characterization of soluble TRAIL, expressed as inclusion bodies in E. coli. sTRAIL inclusion bodies were solubilized and refolded at a high concentration up to 0.9 g/L by a simple dilution method. Refolded protein was purified to electrophoretic homogeneity by a single-step immobilized metal affinity chromatography. The purified sTRAIL had a strong cytotoxic activity against human pancreatic tumor cell line 1990, with ED50 about 1.5 mg/L. Circular dichroism and fluorescence spectrum analysis showed that the refolded sTRAIL had a structure similar to that of native protein with beta-sheet secondary structure. This efficient procedure of sTRAIL renaturation may be useful for the mass production of this therapeutically important protein.