腺病毒蛋白酶：氨基酸序列比较及克隆的cDNA在大肠杆菌中的表达

Adenovirus proteinases: comparison of amino acid sequences and expression of the cloned cDNA in Escherichia coli.

作者信息

Houde A, Weber J M

机构信息

Department of Microbiology, Faculty of Medicine, University of Sherbrooke, Québec, Canada.

出版信息

Gene. 1990 Apr 16;88(2):269-73. doi: 10.1016/0378-1119(90)90042-p.

DOI:10.1016/0378-1119(90)90042-p

PMID:2189788

Abstract

Adenoviruses (Ad) synthesize serine-center endoproteinases (AdEPs) responsible for maturation cleavages within the virus particle. Many questions regarding these enzymes remain unanswered because previous studies utilized crude cells or viral lysates as the enzyme source. Here, we report on the comparison of the amino acid (aa) sequences of several AdEPs and on the expression of the cDNA of the Ad2Ep in Escherichia coli. The AdEPs consist of about 200 aa and their size is around 23 kDa. Among the seven sequences known, 60% of aa were strictly conserved. The usual serine proteinase active site sequence, GDSGG, is absent. The recombinant Ad2EP, produced by an inducible vector as a protein-A fusion product is capable of autocatalytic cleavage, and of cleaving its natural viral substrates as well as foreign proteins. Therefore, other viral proteins or mammalian specific post-translational modifications are not required for enzyme activity.

摘要

腺病毒（Ad）合成负责病毒颗粒内成熟切割的丝氨酸中心内蛋白酶（AdEPs）。由于先前的研究使用粗细胞或病毒裂解物作为酶源，关于这些酶的许多问题仍未得到解答。在此，我们报告了几种AdEPs氨基酸（aa）序列的比较以及Ad2Ep的cDNA在大肠杆菌中的表达。AdEPs由约200个氨基酸组成，其大小约为23 kDa。在已知的七个序列中，60%的氨基酸是严格保守的。通常的丝氨酸蛋白酶活性位点序列GDSGG不存在。由诱导型载体作为蛋白A融合产物产生的重组Ad2EP能够进行自催化切割，并能够切割其天然病毒底物以及外源蛋白质。因此，酶活性不需要其他病毒蛋白或哺乳动物特有的翻译后修饰。