Escherichia coli maltose-binding protein is uncommonly effective at promoting the solubility of polypeptides to which it is fused
Escherichia coli maltose-binding protein is uncommonly effective at promoting the solubility of polypeptides to which it is fused
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DOI：
10.1110/ps.8.8.1668
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发表时间：
1999-08-01
期刊：
影响因子：
8
通讯作者：
Waugh, DS
中科院分区：
文献类型：
--
作者：
Kapust, RB;Waugh, DS
虽然通常有可能在大肠杆菌中获得有利的重组蛋白产量，但以可溶的、生物活性的形式获得蛋白质仍然是一个主要挑战。有时，这个问题可以通过将易于聚集的多肽与高度可溶的伙伴融合来克服。为了更详细地研究这一现象，我们比较了三种可溶性融合伙伴的能力：麦芽糖结合蛋白（MBP）、谷胱甘肽s转移酶；（GST）和硫氧还蛋白（TRX）来抑制六种不同蛋白质的聚集，这些蛋白质通常以不溶性的形式聚集。值得注意的是，我们发现MEP是一种比其他两个融合伙伴更有效的增溶剂。此外。我们证明，在某些情况下，与MEP融合可以促进附着蛋白正确折叠成其生物活性构象。因此，MBP似乎能够在融合蛋白的背景下作为一般分子伴侣发挥作用。提出了一个模型来解释MBP如何促进其溶解度并影响其融合伙伴的折叠。
Although it is usually possible to achieve a favorable yield of a recombinant protein in Escherichia coli, obtaining the protein in a soluble, biologically active form continues to be a major challenge. Sometimes this problem can be overcome by fusing an aggregation-prone polypeptide to a highly soluble partner. To study this phenomenon in greater detail, we compared the ability of three soluble fusion partners-maltose-binding protein (MBP), glutathione S-transferase; (GST), and thioredoxin (TRX)-to inhibit the aggregation of six diverse proteins that normally accumulate in an insoluble form. Remarkably, we found that MEP is a far more effective solubilizing agent than the other two fusion partners. Moreover. we demonstrated that in some cases fusion to MEP can promote the proper folding of the attached protein into its biologically active conformation. Thus, MBP seems to be capable of functioning as a general molecular chaperone in the context of a fusion protein. A model is proposed to explain how MBP promotes the solubility and influences the folding of its fusion partners.