Inhibition of the Membrane Fusion Proteins of Flaviviruses and Alphaviruses

黄病毒和甲病毒膜融合蛋白的抑制

• 批准号: 7255226
• 负责人: MARGARET KIELIAN
• 金额: $ 24.9万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7255226
• 关键词: Alphavirus; Animals; Antiviral Agents; Antiviral Therapy; Binding; Biological Assay; Categories; Cells; Chimeric Proteins; Class; Co-Immunoprecipitations; Collaborations; Complication; Country; Culicidae; Dengue; Dengue Hemorrhagic Fever; Dengue Virus; Developed Countries; Developing Countries; Dominant-Negative Mutation; Encephalitis; Flavivirus; Goals; Health; Helix (Snails); Human; In Vitro; Infection; Institution; Japanese Encephalitis; Lead; Mediating; Membrane; Membrane Fusion; Methods; Molecular Conformation; Numbers; Pathway interactions; Peptide Library; Peptides; Protein Fragment; Proteins; Reaction; Research; Screening procedure; Semliki forest virus; Structure; Testing; Ticks; Transmembrane Domain; United States National Institutes of Health; Viral Hemorrhagic Fevers; Virus; Virus Diseases; Virus Inhibitors; West Nile virus; Western Equine Encephalitis Virus; Yellow fever virus; base; biodefense; dimer; ear helix; high throughput screening; in vitro Assay; inhibitor/antagonist; member; novel; pathogen; protein protein interaction; small molecule; small molecule libraries; tool; trimer core; vector

DESCRIPTION (provided by applicant): Flaviviruses and alphaviruses are spread by mosquito and tick vectors and cause severe human and animal illnesses such as encephalitis and hemorrhagic fever. These viruses include many potential bioterrorist agents that are category A-C pathogens, such as the flaviviruses dengue, West Nile, Japanese encephalitis and yellow fever viruses, and the alphaviruses Venezuelan, eastern, and western equine encephalitis viruses. Dengue virus (DV) is currently of particular concern as it has dramatically reemerged to become endemic in more than 100 countries including the US, and is now a global health problem. There are an estimated 50-100 million cases of dengue fever and 500,000 cases of the more lethal complication dengue hemorrhagic fever per year, with significant impact on both human health and the economies of developing countries. Antiviral strategies for the flaviviruses and alphaviruses are urgently needed. The flavivirus and alphavirus membrane fusion proteins are members of the class II virus fusion proteins. They are structurally very similar and refold to a homotrimer form to mediate virus fusion and infection. In collaboration with Dr. F¿lix Rey, we have recently determined the structure of the homotrimer conformation of the fusion protein from the alphavirus Semliki Forest virus (SFV). The SFV homotrimer structure is strikingly similar to that of DV. Using the structure as a guide, we have developed protein fragments that act as specific dominant-negative inhibitors of SFV and DV fusion and infection. Based on this information, we now plan to develop a general screen for inhibitors of class II fusion reactions. In aim 1 we will establish and optimize in vitro methods to follow the protein-protein interactions during the class II fusion protein refolding reaction. In aim 2, we will adapt this assay to a high throughput format, and use it to screen peptide and small molecule libraries available at our institution and through the NIH and Northeast Biodefense Consortium. Ultimately, such inhibitors will be lead compounds for antiviral therapy, and important research tools to understand the class II virus fusion reaction. Flaviviruses and alphaviruses include many important human pathogens and potential bioterrorist threats such as the flaviviruses dengue, West Nile, Japanese encephalitis and yellow fever viruses, and the alphaviruses Venezuelan, eastern, and western equine encephalitis viruses. Dengue virus is currently of particular concern as it has dramatically reemerged to become endemic in more than 100 countries including the US, and is now a global health problem. This application focuses on developing new antiviral strategies for the flaviviruses and alphaviruses, based on blocking the activity of the proteins involved in the initial entry of the virus into the cell.

描述（由适用提供）：黄病毒和α病毒由蚊子和壁虱矢量传播，并引起严重的人类和动物疾病，例如脑炎和出血热。这些病毒包括许多潜在的生物恐怖剂，它们是A-C类病原体的许多潜在的生物恐怖剂，例如Flavivivires Fans，West Nile，日本脑炎和黄热病病毒，以及委内瑞拉，东部和西马脑炎病毒的α病毒。登革热病毒（DV）目前特别关注，因为它已大大重新出现在包括美国在内的100多个国家 /地区的内构中，现在是一个全球健康问题。估计有500亿例登革热病例，每年有500,000例致命的并发症风扇出血性发烧，对人类健康和发展中国家的经济产生重大影响。迫切需要使用黄病毒和α病毒的抗病毒策略。黄病毒和α病毒膜融合蛋白是II类病毒融合蛋白的成员。它们在结构上非常相似，并将其重新折叠为同构二聚体形式以介导病毒融合和感染。通过与f lix Rey博士的合作，我们最近确定了来自alphavirus semliki森林病毒（SFV）的融合蛋白同构构构象的结构。 SFV同二聚体结构与DV的结构非常相似。使用该结构作为指导，我们开发了蛋白质片段，可作为SFV和DV融合和感染的特定显性阴性抑制剂。基于这些信息，我们现在计划开发一个抑制II类融合反应的通用屏幕。在AIM 1中，我们将建立并优化体外方法，以遵循II类融合蛋白重新折叠反应期间蛋白质蛋白质相互作用。在AIM 2中，我们将将该测定法调整为高通量格式，并使用它来筛选我们机构以及NIH和NORK BIODEFENSE财团可用的肽和小分子库。最终，这种抑制剂将是用于抗病毒疗法的铅化合物，以及了解II类病毒融合反应的重要研究工具。黄酮病毒和α病毒包括许多重要的人类病原体和潜在的生物恐怖主义威胁，例如Flaviviruses风扇，西尼罗河，日本脑炎和黄热病病毒，以及委内瑞拉，东部和西部马脑炎病毒。登革热病毒目前特别关注，因为它已大大重新出现在包括美国在内的100多个国家中的内在，现在是一个全球健康问题。该应用的重点是开发针对黄病毒和α病毒的新抗病毒策略，以阻止病毒初始进入细胞的蛋白质的活性。