RELEASE OF PROTEINS AND PEPTIDES FROM FUSION PROTEINS USING A RECOMBINANT PLANT-VIRUS PROTEINASE
RELEASE OF PROTEINS AND PEPTIDES FROM FUSION PROTEINS USING A RECOMBINANT PLANT-VIRUS PROTEINASE
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DOI：
10.1006/abio.1994.1060
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发表时间：
1994-02-01
影响因子：
2.9
通讯作者：
DOUGHERTY, WG
中科院分区：
文献类型：
--
作者：
PARKS, TD;LEUTHER, KK;DOUGHERTY, WG
描述了一种用于融合蛋白和多肽的生产、切割和纯化的改进方法。这种方法的独特之处在于使用了一种来自烟草蚀刻病毒（TEV）的蛋白酶。所使用的蛋白酶是一种重组TEV蛋白酶，由位于氨基末端的多组氨酸束产生。蛋白酶识别一个特定的，延长的切割位点序列。目的肽或蛋白被纯化为融合蛋白，其TEV蛋白酶裂解位点序列位于其与融合的亲和载体部分之间。用重组TEV蛋白酶孵育介导感兴趣的肽或蛋白质的释放。使用重组TEV蛋白酶来切割融合蛋白是对使用其他蛋白酶的改进，原因包括其高度特异性，对通常用于蛋白质纯化的许多蛋白酶抑制剂不敏感，并且亲和标签和蛋白酶都可以从感兴趣的切割产物中分离出来。
An improved method for the production, cleavage, and purification of fusion proteins and peptides is described. The unique aspect of this method is dependent on the use of a proteinase from tobacco etch virus (TEV). The proteinase used is a recombinant TEV proteinase produced with a polyhistidine tract positioned at the amino terminus. The proteinase recognizes a specific, extended cleavage site sequence. The peptide or protein of interest is purified as a fusion protein with a TEV proteinase cleavage site sequence located between it and an affinity carrier portion of the fusion. Incubation with the recombinant TEV proteinase mediates release of the peptide or protein of interest. Use of the recombinant TEV proteinase to cleave fusion proteins is an improvement over use of other proteinases for several reasons, including its high degree of specificity, its insensitivity to many proteinase inhibitors generally used in protein purification, and the ready separation of both the affinity tag and the proteinase from the cleaved product of interest.