CHARACTERIZATION OF CMV PROTEASE IN CAPSID ASSEMBLY

衣壳组装中 CMV 蛋白酶的表征

• 批准号: 6215469
• 负责人: BARRY C HOLWERDA
• 金额: $ 22.43万
• 依托单位: UNIVERSITY OF MISSISSIPPI MED CTR
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6215469
• 关键词: cytomegalovirus; endopeptidases; enzyme activity; gene mutation; laboratory mouse; protease inhibitor; protein structure function; recombinant virus; virus assembly; virus protein

The object of this proposal is to further characterize cytomegalovirus protease and in so doing to aid in the development of antiviral inhibitors. The central hypothesis is that because protease function is intimately associated with capsid structure and assembly, studying the catalytic properties of recombinant protease does not allow prediction of the antiviral effects of protease inhibitors to be made with any accuracy. This application proposes to construct strains of murine cytomegalovirus (MCMV) in which the protease/assembly gene has been mutated and to characterize those mutants with respect to a variety of properties, including replication, morphology, and protein-protein interactions in the virion capsid. A second part of the proposal is to purify native MCMV protease and compare it with recombinant protease to try to obtain an enzyme that can be used to more accurately study the mechanisms of action of PR antivirals. A chimeric MCMV containing a human CMV (HCMV) protease will be constructed and tested for the ability to grow in mice. If successful, this will provide a way to test HCMV antivirals in vivo and to correlate the results with those from analyses of recombinant and native enzymes.

这项提议目的是进一步描述 巨细胞病毒蛋白水解酶及其在抗病毒药物开发中的作用 抑制剂。中心假设是，因为蛋白酶的功能是 与衣壳结构和组装密切相关，研究 重组酶的催化特性不允许预测 蛋白水解酶抑制剂的抗病毒作用要精确到哪一步。这 小鼠巨细胞病毒(MCMV)株在中国的应用 哪些蛋白水解酶/组装基因发生了突变，并对这些基因进行了表征 突变体涉及各种属性，包括复制， 病毒粒子衣壳的形态和蛋白质-蛋白质的相互作用。一秒钟 该提案的一部分是纯化天然的MCMV蛋白酶，并与 重组蛋白酶试图获得一种可用于更多的酶 准确研究PR抗病毒药物的作用机制。一种嵌合的小细胞巨细胞病毒 将构建包含人巨细胞病毒(HCMV)蛋白酶的 在老鼠体内生长的能力。如果成功，这将提供一种检测HCMV的方法 体内的抗病毒药物，并将结果与分析的结果相关联 重组酶和天然酶。