意想不到的烟草蚀纹病毒（TEV）蛋白酶对重组人蛋白的切割。
Unexpected tobacco etch virus (TEV) protease cleavage of recombinant human proteins.
机构信息
Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.
Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.
出版信息
Protein Expr Purif. 2024 Aug;220:106488. doi: 10.1016/j.pep.2024.106488. Epub 2024 Apr 26.
The tobacco etch virus (TEV) protease is a commonly used reagent for removal of solubility and purification tags from recombinant proteins and is cited as being highly specific for its canonical cleavage site. Flexibility in some amino acids within this recognition sequence has been described in the literature but researchers generally assume few native human proteins will carry off-target sequences for TEV cleavage. We report here the aberrant cleavage of three human proteins with non-canonical TEV protease cleavage sites and identify broader sequence specificity rules that can be used to predict unwanted cleavage of recombinant proteins. Using these rules, 456 human proteins were identified that could be substrates for unwanted TEV protease cleavage.
烟草蚀纹病毒（TEV）蛋白酶是一种常用于从重组蛋白中去除可溶性和纯化标签的试剂，被认为对其典型的切割位点具有高度特异性。文献中描述了该识别序列中一些氨基酸的灵活性，但研究人员通常认为很少有天然存在的人类蛋白会带有 TEV 切割的非靶序列。我们在此报告三个具有非典型 TEV 蛋白酶切割位点的人类蛋白的异常切割，并确定了更广泛的序列特异性规则，可用于预测重组蛋白的非预期切割。使用这些规则，鉴定出 456 个人类蛋白可能是 TEV 蛋白酶非预期切割的底物。
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