烟草蚀刻病毒（TEV）蛋白酶意外裂解重组人类蛋白质

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2024-04-26 DOI:10.1016/j.pep.2024.106488

Lauren P. Beaumont, Jennifer Mehalko, Adam Johnson, Vanessa E. Wall, Dominic Esposito

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引用次数: 0

摘要

烟草蚀刻病毒（TEV）蛋白酶是一种常用的试剂，用于去除重组蛋白中的溶解性和纯化标签，并被认为对其典型裂解位点具有高度特异性。文献中描述了该识别序列中某些氨基酸的灵活性，但研究人员普遍认为很少有原生人类蛋白质会携带 TEV 裂解的非目标序列。我们在此报告了三种带有非典型 TEV 蛋白酶裂解位点的人类蛋白质的异常裂解，并确定了更广泛的序列特异性规则，这些规则可用于预测重组蛋白质的意外裂解。利用这些规则，共鉴定出 456 种可能成为 TEV 蛋白酶意外裂解底物的人类蛋白质。

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Unexpected tobacco etch virus (TEV) protease cleavage of recombinant human proteins

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.

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.