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：疱疹组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 该蛋白质的巨细胞病毒（CMV）同源物称为组装细胞软管小组，为 由人类CMV中的UL80A开放阅读框编码。 汇编为 合成为前体（〜74KDA），该前体经历四个顺序 自溶解分解。 这四个切割部位有五个氨基酸， 核心共识序列在其对应物中充分保守 其他疱疹病毒。 组件的主要基材是丰富的 衣壳组件蛋白也包含其M-Cleavage位点 羧基端，由于其有趣的框架嵌套遗传 与蛋白酶的关系。 因为这些分裂对于病毒的产生至关重要，所以 蛋白酶有望具有有效的抗病毒作用。 新的 需要针对疱疹病毒的抗病毒活性的药物，并且 本应用中提出的研究旨在进一步 表征了物理，酶和生物学特性 疱疹病毒蛋白酶，尤其是CMV酶，并将其剥削为 药物开发的有效分子靶标。 在此竞争更新中提出的工作的具体目的 应用是：（i）准备蛋白酶和装配蛋白前体 用于酶测定和晶体学； ii）发展体外蛋白酶 使用“本机”底物'（iii）确定哪种裂解位点的测定 特征会影响切割动力学； （iv）识别交互式域 两个链条组件； （v）合作研究和比较伽马2 HHV8 带有其α（例如，单纯疱疹病毒）和β（例如CMV）的alpha（例如，CMV） 疱疹病毒同源物； （vi）测试细胞培养中潜在的抗病毒药；和 （vii）确定病毒“逃脱”其作用的能力。 预计这项工作的结果将提供有关有关的有用新信息 这个显然是丝氨酸蛋白酶家族的新成员，并有助于 开发将阻止其功能的抑制剂。 也是 病毒区域可能会生成有用的新信息 蛋白酶机制和疱疹病毒复制。