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马铃薯Y病毒辅助成分蛋白酶在大肠杆菌中及体外的自催化加工

Autocatalytic processing of the potyvirus helper component proteinase in Escherichia coli and in vitro.

作者信息

Carrington J C, Freed D D, Sanders T C

机构信息

Department of Biology, Texas A&M University, College Station 77843-3258.

出版信息

J Virol. 1989 Oct;63(10):4459-63. doi: 10.1128/JVI.63.10.4459-4463.1989.

DOI:10.1128/JVI.63.10.4459-4463.1989
PMID:2674480
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC251070/
Abstract

The virus-encoded proteins of tobacco etch virus (TEV), a plant potyvirus, arise by proteolytic processing of a large polyprotein precursor. The TEV genome codes for two proteinases, a 49-kilodalton proteinase and helper component proteinase (HC-Pro), which cleave the polyprotein at specific sites. The only known cleavage event catalyzed by HC-Pro occurs at the HC-Pro carboxyl terminus. The proteolytic activity of HC-Pro was analyzed by expression of the enzyme in bacterial and cell-free systems. The carboxyl-terminal domain of HC-Pro exhibited proteolytic activity in Escherichia coli with a processing half-time of approximately 100 s. The processing kinetics of HC-Pro expressed in vitro by cell-free transcription and translation was variable, depending on the presence or absence of TEV polypeptide sequences at the amino terminus of the proteolytic domain. Cleavage of the HC-Pro carboxyl terminus appeared to proceed exclusively by an autocatalytic mechanism; the proteinase synthesized in vitro exhibited little or no proteolytic activity when reacted with the HC-Pro cleavage site in trans or biomolecular reactions.

摘要

烟草蚀纹病毒（TEV）是一种植物马铃薯Y病毒属病毒，其病毒编码蛋白由一个大的多蛋白前体经蛋白水解加工产生。TEV基因组编码两种蛋白酶，一种49千道尔顿的蛋白酶和辅助成分蛋白酶（HC-Pro），它们在特定位点切割多蛋白。已知由HC-Pro催化的唯一切割事件发生在HC-Pro的羧基末端。通过在细菌和无细胞系统中表达该酶来分析HC-Pro的蛋白水解活性。HC-Pro的羧基末端结构域在大肠杆菌中表现出蛋白水解活性，加工半衰期约为100秒。通过无细胞转录和翻译在体外表达的HC-Pro的加工动力学是可变的，这取决于蛋白水解结构域氨基末端是否存在TEV多肽序列。HC-Pro羧基末端的切割似乎完全通过自催化机制进行；体外合成的蛋白酶在反式或双分子反应中与HC-Pro切割位点反应时，表现出很少或没有蛋白水解活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/251070/ab661ec38317/jvirol00077-0354-b.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/251070/e525c1908e39/jvirol00077-0354-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/251070/ab661ec38317/jvirol00077-0354-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/251070/352264dfc9ea/jvirol00077-0352-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/251070/8bbac528b71a/jvirol00077-0353-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/251070/e525c1908e39/jvirol00077-0354-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bd3/251070/ab661ec38317/jvirol00077-0354-b.jpg

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