Mocr: A novel fusion tag for enhancing solubility that is compatible with structural biology applications
Mocr: A novel fusion tag for enhancing solubility that is compatible with structural biology applications
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DOI：
10.1016/j.pep.2008.08.011
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发表时间：
2009-01-01
影响因子：
1.6
通讯作者：
Brown, William Clay
中科院分区：
文献类型：
--
作者：
DelProposto, James;Majmudar, Chinmay Y.;Brown, William Clay
异源蛋白生产中的一个持续问题是当在宿主细胞中高水平表达时靶蛋白的不溶性。克服该问题的广泛采用的策略是使用融合标签。最好的融合标签促进溶解度。可以起到纯化手柄的作用，并且不干扰下游应用或者可以从过客蛋白制剂中除去。鉴定出满足这些标准的新型融合标签。该融合标签是噬菌体T7的Ocr蛋白（0.3基因产物）的单体突变体。该融合标签显示出与多种不同乘客蛋白的增溶活性。我们表明，它可以用作类似于其他融合标签的纯化手柄。它的小尺寸和紧凑结构与其在过客蛋白的下游应用中的用途相容，或者它可以从过客蛋白中除去和纯化。使用单体Ocr（Mocr）作为其他融合标签如麦芽糖结合蛋白的互补物将在蛋白质生产和加工中提供更大的灵活性，用于各种蛋白质应用。(c)2008年爱思唯尔公司All rights reserved.
A persistent problem in heterologous protein production is insolubility of the target protein when expressed to high level in the host cell, A widely employed strategy for overcoming this problem is the use of fusion tags. The best fusion tags promote solubility. may function as purification handles and either do not interfere with downstream applications or may be removed from the passenger protein preparation. A novel fusion tag is identified that meets these criteria. This fusion tag is a monomeric mutant of the Ocr protein (0.3 gene product) of bacteriophage T7. This fusion tag displays solubilizing activity with a variety of different passenger proteins. We show that it may be used as a purification handle similar to other fusion tags. Its small size and compact structure are compatible with its use in downstream applications of the passenger protein or it may be removed and purified away from the passenger protein. The use of monomeric Ocr (Mocr) as a complement to other fusion tags such as maltose-binding protein will provide greater flexibility in protein production and processing for a wide variety of protein applications. (c) 2008 Elsevier Inc. All rights reserved.