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.

疱疹病毒成熟蛋白酶对于感染性病毒的产生至关重要，并且是合成为在衣壳组装过程中具有多种功能的前体。自发现以来 12 多年前，它作为抗病毒药物的可能目标而受到相当大的关注，并且在过去 7 年里关于这种有趣但具有挑战性的酶的大量信息已经积累起来。 28 kDa 该酶是通过自蛋白水解裂解从 74 kDa 前体衍生而来，首先在 (M) 饱和位点靠近其羧基末端，然后在（R）释放位点。 R 位点裂解将装配蛋白从 ---44-kDa 中释放出来前体的羧基部分。蛋白水解结构域，在巨细胞病毒中称为装配蛋白（Assemblin），已被克隆、纯化和体外研究。该酶通过二聚化被激活，但二聚体对有两个此外，它与其他丝氨酸蛋白酶的新折叠（7- 链[3桶]和催化三联体（Ser-His-His）。没有关于该产品的可比信息前驱。我们的目标是通过生化和遗传学研究更多地了解这种病毒酶，特别关注其前身。具体目标旨在通过以下方式帮助实现该目标： (i) 应用突变病毒来研究病毒感染过程中该蛋白酶的机制，(ii) 开发一种蛋白酶突变体，其活性可以通过化学救援在细胞内和体外开启，(iii) 研究两个催化位点突变体形成选择性互补复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学性质与组装蛋白的酶学性质进行比较，以及（v）研究两种形式的酶二聚化的要求和后果。结果这项工作预计将提供有关丝氨酸这一新成员的有用的新信息蛋白酶家族，并有助于开发阻断其功能的抑制剂。也有可能在病毒蛋白酶机制和疱疹病毒领域将产生有用的新信息复制。

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。