Expression, purification, and renaturation of bone morphogenetic protein-2 from Escherichia coli
Expression, purification, and renaturation of bone morphogenetic protein-2 from Escherichia coli
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DOI：
10.1016/j.pep.2005.09.025
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发表时间：
2006-04-01
影响因子：
1.6
通讯作者：
Ma, HW
中科院分区：
文献类型：
--
作者：
Long, SN;Truong, L;Ma, HW
同源二聚体骨形态发生蛋白-2（BMP-2）是转化生长因子β超家族的成员，已用于骨移植。本研究旨在探索BMP-2在其他疾病中的功能，并致力于表达和纯化具有活性的BMP-2蛋白。我们已经开发了一种新的方法，该方法涉及使用FoldIt重折叠缓冲液重折叠BMP-2，然后使用肝素亲和柱将正确折叠的二聚体与单体分离。获得了每克细胞湿重29.4mg BMP-2二聚体的高产率。通过在C2 C12细胞中诱导碱性磷酸酶活性测试，纯化的BMP-2二聚体显示具有与来自CHO细胞的BMP-2相同的活性水平。这种方法在复性和纯化其他同源二聚体蛋白质方面具有潜在的应用价值。(c)2005年爱思唯尔公司All rights reserved.
Homodimeric bone morphogenetic protein-2 (BMP-2) is a member of the transforming growth factor beta superfamily that has been used for bone grafting. We were interested in exploring the functions of BMP-2 in other disease areas and focused on expressing and purifying active BMP-2 proteins. We have developed a new approach which involves using FoldIt refolding buffer to refold BMP-2 followed by a heparin affinity column to separate correctly folded dimer from monomer. A high yield of 29.4 mg BMP-2 dimer per gram cell wet weight was achieved. The purified BMP-2 dimer was shown to possess the same level of activity as BMP-2 from CHO cells as tested by the induction of alkaline phosphatase activity in C2C12 cells. This approach has potential application in refolding and purifying other homodimeric proteins. (c) 2005 Elsevier Inc. All rights reserved.