Denque Virus Receptor Interactions

描述（由申请人提供）：登革热病毒感染的复苏 是热带发展中国家的主要公共卫生威胁;没有 有效的治疗或疫苗。我们研究的主要假设是 登革病毒包膜蛋白与 靶细胞上的受体代表了致病性的关键机制，并且 理解这种相互作用将使我们能够有效地 基于抑制病毒-靶细胞结合的治疗策略。的 项目包括三个互补的目标;初步数据包括 三是实现三个目标的可行性。一种独特的硫酸乙酰肝素 蛋白聚糖是一种保守的登革病毒受体， 来自多个器官和物种。糖胺聚糖的成分 蛋白聚糖体现了病毒包膜蛋白结合活性。主 目的1的目标是使用包膜来充分表征这种糖胺聚糖， 蛋白质亲和层析，然后解聚， 寡糖测序。我们还将尝试鉴定蛋白聚糖 核心蛋白 登革病毒包膜蛋白负责细胞结合活性。目标 目的2是鉴定特异性包膜蛋白区域和氨基酸 负责细胞结合。我们将使用定点突变来产生 突变的包膜蛋白，并确定突变导致的损失， 约束力为了证实突变的生物学意义，重组 将产生包含突变的病毒，并评估其损失。 细胞结合为了证实（初始数据）来自临床的包膜蛋白 分离株与实验室登革热病毒株结合相同的受体，我们将 还比较了来自非传代登革热的包膜蛋白的结合谱， 病毒临床分离物与实验室病毒衍生的包膜蛋白。 可溶性糖胺聚糖和聚磺酸苏拉明预防登革热病毒 包膜蛋白结合靶细胞，并防止感染;化学 过硫酸化糖胺聚糖是特别有效的抑制剂。的目标 目的3是合成硫酸化/唾液酸化分子，基于这些分子， 并测试它们在抑制包膜蛋白结合和登革热中的活性 病毒传染性

DESCRIPTION (provided by applicant): The resurgence of dengue virus infection is a major public health threat to the tropical developing world; there is no effective treatment or vaccine. The primary hypothesis underlying our research program is that interactions between dengue virus envelope protein and receptors on target cells represents a pivotal mechanism of pathogenicity, and that understanding this interaction will enable us to develop effective therapeutic strategies based on inhibition of virus-target cell binding. The project includes three complementary aims; preliminary data are included to document the feasibility of all 3 aims. A distinctive heparan sulfate proteoglycan is a conserved dengue virus receptor expressed by cells derived from multiple organs and species. The glycosaminoglycan component of the proteoglycan embodies the virus envelope protein binding activity. The primary goal of Aim 1 is to fully characterize this glycosaminoglycan, using envelope protein affinity chromatography, followed by depolymerization and oligosaccharide sequencing. We will also attempt to identify the proteoglycan core protein. The dengue virus envelope protein accounts for cell-binding activity. The goal of Aim 2 is to identify the specific envelope protein region and amino acids responsible for cell-binding. We will use site-directed mutagenesis to generate mutant envelope proteins, and identify mutations that result in loss of binding. To confirm the biological significance of mutations, recombinant viruses incorporating mutations will be generated, and assessed for loss of cell binding. To confirm (initial data) that envelope proteins from clinical isolates bind the same receptor as laboratory dengue virus strains, we will also compare the binding profiles of envelope proteins from non-passaged dengue virus clinical isolates with laboratory-virus derived envelope proteins. Soluble glycosaminoglycans, and the polysulfonate Suramin prevent dengue virus envelope protein binding to target cells, and prevent infection; chemically persulfated glycosaminoglycans are particularly potent inhibitors. The goal of Aim 3 is to synthesize sulfated/sialylated molecules, based on these molecules, and to test their activity in inhibiting envelope protein binding and dengue virus infectivity.