Herpesvirus Proteinase - Possible Target for Antivirals

疱疹病毒蛋白酶——抗病毒药物的可能靶点

基本信息

批准号：
7156940
负责人：
D Wade Gibson
金额：
$ 27.13万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1992
资助国家：
美国
起止时间：
1992-07-01 至 2008-12-31
项目状态：
已结题

项目摘要

The herpesvirus maturational protease is essential for the production of infectious virus and is synthesized as a precursor that has multiple functions during capsid assembly. Since its discovery 12 years ago, it has received considerable attention as a possible target for antivirals, and in the past 7 years a wealth of information about this interesting, albeit challenging, enzyme has accumulated. The 28-kDa enzyme is derived from a 74-kDa precursor by autoproteolytic cleavage, first at the (M)aturational site near its carboxyl end, and then at the (R)elease site. R-site cleavage frees assemblin from the ---44-kDa carboxyl portion of the precursor. The proteolytic domain, called assemblin in cytomegaolvirus, has been cloned, purified, and studied in vitro. The enzyme is activated by dimerization but the dimer pair has two separate acitive sites, moreover, it differs remarkably from other serine proteases by its new fold (7- stranded [3 barrel) and catalytic triad (Ser-His-His). No comparable information is available about the precursor. Our objective is to learn more about this viral enzyme through biochemical and genetic studies, with particular attention to its precursor. The specific aims are intended to help accomplish that objective by (i) applying mutant viruses to study the mechanism of this protease during virus infection, (ii) developing a protease mutant whose activity can be switched on in cells and in vitro by chemical rescue, (iii) investigating the ability of two catalytic-site mutants to form a complementation complex with selective specificity for the maturational-cleavage site, (iv) taking advantage of recent findings to investigate the structure of the precursor and compare its enzymatic properties with those of assemblin, and (v) investigating requirements and consequences of dimerization by both forms of the enzyme. Results of this work are anticipated to provide useful new information about this novel member of the serine proteinase family, and contribute to development of inhibitors that will block its function. It is also likely that useful new information will be generated in the areas of viral proteinase mechanisms and herpesvirus replication.

疱疹病毒的成熟蛋白酶对于产生传染病至关重要，是合成为在衣壳组装过程中具有多个功能的前体。自发现12 几年前，它已被广泛关注，作为抗病毒药的可能目标，在过去的7年中关于这种有趣但充满挑战的酶的大量信息已经积累。 28 kda 酶是源自自动溶解裂解的74 kDa前体，首先是在（M）占地位点在其羧基端附近，然后在（R）省略地点。 R-Site CLEAVAGE FREES FREES Assemblin从 - 44 kDa 前体的羧基部分。蛋白水解结构域，称为cytomegaolvirus中的汇编小组，已经是克隆，纯化和研究体外研究。酶通过二聚化激活，但二聚体对有两个此外，单独的阳性位点与其他丝氨酸蛋白酶的新折叠明显不同（7-- 滞留[3桶）和催化三合会（ser-His-His）。没有关于前体。我们的目标是通过生化和遗传研究更多地了解这种病毒酶特别注意其前体。具体目标旨在帮助实现这一目标（i）应用突变病毒在病毒感染过程中研究这种蛋白酶的机制，（ii）一个蛋白酶突变体，其活性可以在细胞中打开并通过化学救援体外打开，（iii）研究两个催化位点突变体形成具有选择性的互补配合物的能力成熟 - 裂解位点的特异性，（iv）利用最近的发现来研究前体的结构，并将其酶特性与总结细胞的结构进行比较，（V）通过两种形式的酶调查二聚化的要求和后果。结果预计这项工作将提供有关丝氨酸这一小说成员的有用新信息蛋白酶家族，并有助于阻断其功能的抑制剂的发展。也可能该有用的新信息将在病毒蛋白酶机制和疱疹病毒的领域产生复制。

项目成果

期刊论文数量（17）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Structure and formation of the cytomegalovirus virion.

DOI：
10.1007/978-3-540-77349-8_11
发表时间：
2008
期刊：
Current topics in microbiology and immunology
影响因子：
0
作者：
Wade Gibson
通讯作者：
Wade Gibson

Independently cloned halves of cytomegalovirus assemblin, An and Ac, can restore proteolytic activity to assemblin mutants by intermolecular complementation.

独立克隆的巨细胞病毒装配蛋白 An 和 Ac 的一半可以通过分子间互补恢复装配蛋白突变体的蛋白水解活性。

DOI：
10.1128/jvi.71.2.956-964.1997
发表时间：
1997
期刊：
Journal of virology.
影响因子：
0
作者：
Hall,MR;Gibson,W
通讯作者：
Gibson,W

Chemical rescue of I-site cleavage in living cells and in vitro discriminates between the cytomegalovirus protease, assemblin, and its precursor, pUL80a.

活细胞和体外 I 位点裂解的化学拯救可区分巨细胞病毒蛋白酶、组装蛋白及其前体 pUL80a。

DOI：
10.1074/jbc.m506876200
发表时间：
2005
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
McCartney,StephenA;Brignole,EdwardJ;Kolegraff,KeliN;Loveland,AmyN;Ussin,LaShonM;Gibson,Wade
通讯作者：
Gibson,Wade

Cytomegalovirus assembly protein precursor and proteinase precursor contain two nuclear localization signals that mediate their own nuclear translocation and that of the major capsid protein.

巨细胞病毒组装蛋白前体和蛋白酶前体含有两个核定位信号，介导其自身的核易位和主要衣壳蛋白的核易位。