PICORNAVIRAL PROCESSING PROTEINASES

小RNA病毒加工蛋白酶

• 批准号: 3547944
• 负责人: Ben M. Dunn
• 金额: $ 19.72万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3547944
• 关键词: Hepatovirus; Picornaviridae; X ray crystallography; active sites; aldehydes; antiviral agents; chemical binding; cysteine; drug design /synthesis /production; endopeptidases; enzyme substrate complex; fluorimetry; high performance liquid chromatography; mass spectrometry; peptide chemical synthesis; poliovirus; protease inhibitor; protein structure function; rhinovirus

Our long term objective is to develop potent and selective inhibitors of the picornaviral processing enzymes that may be of value in antiviral therapy. To achieve this goal, we will study the catalytic and binding properties of the 3C cysteine proteinases of hepatitis A virus, human rhinovirus types 2 and 14 and human poliovirus. The collaborating group, led by Bruce Malcolm at Protos, has expressed large quantities of the hepatitis enzyme. We have examined a variety of potential peptide sub- strates and developed rapid, quantitative assays. We are now poised to expand our study by refining the peptide structure that is recognized by this enzyme, by developing new assay methods based on sensitive fluorescence methods, and by constructing potential inhibitors based on peptide structures. We will utilize all these methods to thoroughly analyze the interaction of substrates and inhibitors with the Hepatitis enzyme. As the hepatitis enzyme is in hand, it will serves as our initial target. However, we will pursue the closely related rhinovirus and poliovirus enzymes by the same approach. We anticipate that each system will provide new insights into selective inhibitors design. The human cysteine proteinases found in the lysosomal compartment of most human cells, cathepsins B,H, and L, will be utilized in studies for the specificity of compounds designed against the picornaviral proteinases. Samples of enzymes and first-generation inhibitor will be provided to a collaborating crystallography group. Data arising from this approach will be utilized to design newer inhibiotrs in a further effort to improve the selectivity.

我们的长期目标是开发有效且有选择性的 小核糖核酸病毒加工酶的抑制剂可能具有以下价值 抗病毒治疗。 为了实现这一目标，我们将研究催化和结合 人类甲型肝炎病毒 3C 半胱氨酸蛋白酶的特性 2 型和 14 型鼻病毒以及人类脊髓灰质炎病毒。 合作小组， 由 Protos 的 Bruce Malcolm 领导，已经表达了大量 肝炎酶。 我们已经检查了多种潜在的肽亚 策略并开发了快速定量分析。 我们现在准备 通过完善已被认可的肽结构来扩展我们的研究 这种酶，通过开发基于敏感的新测定方法 荧光方法，并通过构建潜在的抑制剂 肽结构。 我们将利用所有这些方法来彻底 分析底物和抑制剂与肝炎的相互作用 酶。 由于肝炎酶在手，它将作为我们最初的 目标。 然而，我们将追寻密切相关的鼻病毒和 脊髓灰质炎病毒酶采用相同的方法。 我们预计每个系统 将为选择性抑制剂设计提供新的见解。 在溶酶体区室中发现的人类半胱氨酸蛋白酶 大多数人类细胞，组织蛋白酶 B、H 和 L，将用于以下研究： 针对小核糖核酸病毒设计的化合物的特异性 蛋白酶。 将提供酶和第一代抑制剂样品 到一个合作晶体学小组。 由此产生的数据 进一步努力，将利用该方法设计更新的抑制剂 以提高选择性。