HERPESVIRUS PROTEINASE--POSSIBLE TARGET FOR ANTIVIRALS

疱疹病毒蛋白酶——抗病毒药物的可能靶标

• 批准号: 6170156
• 负责人: D Wade Gibson
• 金额: $ 19.94万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 2002-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6170156
• 关键词: Alphaherpesvirinae; Gammaherpesvirinae; Herpesviridae; active sites; antiviral agents; cytomegalovirus; enzyme biosynthesis; enzyme mechanism; enzyme structure; human tissue; protein structure function; proteolysis; serine proteinases; site directed mutagenesis; tissue /cell culture; transfection; virus protein

DESCRIPTION: Herpes-group viruses encode a serine maturational proteinase that is essential for the production of infectious progeny. The cytomegalovirus (CMV) homologue of this protein is called assemblin and is encoded by the UL80a open reading frame in human CMV. Assemblin is synthesized as a precursor (~74kDa) that undergoes four sequential autoproteolytic cleavages. The four cleavage sites have five amino acid, core consensus sequences that are well conserved among their counterparts in other herpes virus. The principal substrate of assemblin is an abundant capsid assembly protein that also contains the M-cleavage site at its carboxyl end, as a consequence of its interesting in-frame, nested genetic relationship with the proteinase. Because these cleavages are essential for virus production, inhibition of the proteinase would be expected to have a potent antiviral effect. New drugs with antiviral activity against herpes-group viruses are needed, and the studies proposed in this application are intended to further characterize the physical, enzymatic, and biological properties of the herpesvirus proteinase, in particular the CMV enzyme, and help exploit it as an effective molecular target for drug development. The specific aims of the work proposed in this competitive renewal application are: (i) prepare proteinase and assembly protein precursors for use in enzyme assays and crystallography; ii) develop an in vitro proteinase assay that uses "native" substrate' (iii) determine what cleavage-site features influence cleavage kinetics; (iv) identify the interactive domains of two chain assemblin; (v) collaborate to study and compare gamma-2 HHV8 assemblin with its alpha (e.g., herpes simplex virus) and beta (e.g., CMV) herpesvirus homologues; (vi) test potential antivirals in cell culture; and (vii) determine the ability of virus to "escape" their effect. Results of this work are anticipated to provide useful new information about this apparently novel member of the serine proteinase family, and contribute to the 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.

描述：疱疹病毒编码丝氨酸成熟蛋白酶 这对于产生传染性后代至关重要。 这 该蛋白的巨细胞病毒 (CMV) 同源物称为装配蛋白，是 由人类 CMV 中的 UL80a 开放阅读框编码。 汇编是 合成为前体 (~74kDa)，经历四个连续的过程 自蛋白水解切割。 四个切割位点有五个氨基酸， 核心共有序列在其对应序列中保存完好 其他疱疹病毒。 组装蛋白的主要底物是丰富的 衣壳组装蛋白，其还含有 M 切割位点 羧基末端，由于其有趣的框内嵌套遗传 与蛋白酶的关系。 因为这些切割对于病毒的产生至关重要，因此抑制 预计该蛋白酶具有有效的抗病毒作用。 新的 需要具有抗疱疹病毒活性的药物，并且 本申请中提出的研究旨在进一步 表征物理、酶学和生物特性 疱疹病毒蛋白酶，特别是 CMV 酶，并帮助利用它作为 药物开发的有效分子靶点。 本次竞争性更新中提出的工作的具体目标 应用是： (i) 制备蛋白酶和组装蛋白前体 用于酶测定和晶体学； ii) 开发体外蛋白酶 使用“天然”底物的测定' (iii) 确定什么切割位点 特征影响裂解动力学； (iv) 确定交互域 两条链条组装； (v) 合作研究和比较 gamma-2 HHV8 装配蛋白及其α（例如单纯疱疹病毒）和β（例如CMV） 疱疹病毒同源物； (vi) 在细胞培养物中测试潜在的抗病毒药物；和 (vii) 确定病毒“逃脱”其影响的能力。 这项工作的结果预计将提供有用的新信息 这显然是丝氨酸蛋白酶家族的新成员，并有助于 开发阻止其功能的抑制剂。 这也是 病毒领域可能会产生有用的新信息 蛋白酶机制和疱疹病毒复制。