Davis Stella M, Romig Bryn L, Abe Alyssa A, Loening Nikolaus M
Department of Chemistry, Lewis & Clark College, Portland, Oregon, USA.
Protein Sci. 2025 Mar;34(3):e70049. doi: 10.1002/pro.70049.
Here we show that a combination of previously suggested mutations for tobacco etch virus (TEV) protease results in a TEV protease mutant that maintains the same catalytic efficiency as previously described mutants but has enhanced stability and solubility. Another advantage of this new variant of TEV protease is that it does not need the inclusion of a reducing agent to maintain its effectiveness, making it easier to generate, store, and use in cleavage reactions compared to previous TEV protease mutants and, in particular, makes it a good choice for cleaving proteins that contain disulfide bonds that would otherwise be altered by the inclusion of a reducing agent. We also provide a straightforward purification protocol for generating this new version of TEV protease.
在此我们表明，烟草蚀纹病毒（TEV）蛋白酶先前提出的多种突变组合产生了一种TEV蛋白酶突变体，该突变体保持了与先前描述的突变体相同的催化效率，但稳定性和溶解性得到了增强。这种新型TEV蛋白酶变体的另一个优点是，它不需要添加还原剂来维持其有效性，与先前的TEV蛋白酶突变体相比，更易于制备、储存并用于切割反应，尤其对于切割含有二硫键的蛋白质而言是个不错的选择，因为加入还原剂会改变这些蛋白质，而该变体则不会。我们还提供了一种简单的纯化方案来制备这种新型TEV蛋白酶。
