A HERPESVIRUS MATURATIONAL PROTEINASE, ASSEMBLIN - IDENTIFICATION OF ITS GENE, PUTATIVE ACTIVE-SITE DOMAIN, AND CLEAVAGE SITE
A HERPESVIRUS MATURATIONAL PROTEINASE, ASSEMBLIN - IDENTIFICATION OF ITS GENE, PUTATIVE ACTIVE-SITE DOMAIN, AND CLEAVAGE SITE
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DOI：
10.1073/pnas.88.23.10792
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发表时间：
1991-12-01
影响因子：
11.1
通讯作者：
GIBSON, W
中科院分区：
文献类型：
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作者：
WELCH, AR;WOODS, AS;GIBSON, W
通过在瞬时转染试验中使用克隆的酶和底物基因，已经鉴定并部分表征了疱疹病毒蛋白酶活性。巨细胞病毒科尔本株的蛋白酶基因编码一个590个氨基酸的蛋白，其N-末端249个残基含有蛋白水解活性和两个结构域，在其他疱疹病毒的同源蛋白中高度保守。在这些结构域之间插入短氨基酸序列会消除蛋白酶活性，这表明该区域构成了酶活性位点的部分或全部。等离子体解吸质谱法用于鉴定成熟组装蛋白的C末端为丙氨酸，使得能够识别所有疱疹病毒组装蛋白同源物的C末端附近的V/L-X-A向下S/V的共有蛋白酶切割序列。有趣的是，蛋白酶和它的底物，即组装蛋白前体，是由同一开放阅读框架的相对两半编码的。
A herpesvirus proteinase activity has been identified and partially characterized by using the cloned enzyme and substrate genes in transient transfection assays. Evidence is presented that the proteinase gene of cytomegalovirus strain Colburn encodes a 590-amino acid protein whose N-terminal 249 residues contain the proteolytic activity and two domains that are highly conserved in the homologous protein of other herpesviruses. Insertion of a short amino acid sequence between these domains abolished proteinase activity, suggesting that this region constitutes part or all of the enzyme active site. Plasma desorption mass spectrometry was used to identify the C terminus of the mature assembly protein as alanine, enabling the recognition of a consensus proteinase cleavage sequence of V/L-X-A down S/V, near the C-terminal end of all herpesvirus assembly protein homologs. Interestingly, the proteinase and its substrate, the assembly protein precursor, are encoded by opposite halves of the same open reading frame.