Interacting motifs of alphavirus envelope proteins.

甲病毒包膜蛋白的相互作用基序。

基本信息

批准号：
6775634
负责人：
WERNER A BRAUN
金额：
$ 30.03万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-08-01 至 2007-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6775634
关键词：
Alphavirus Venezuelan equine encephalitis virus binding sites bioterrorism /chemical warfare computer program /software computer simulation crosslink gene mutation genetic mapping glycoproteins molecular biology information system monoclonal antibody protein protein interaction receptor binding receptor expression virulence virus envelope western blottings

项目摘要

DESCRIPTION (provided by applicant): The goal of this project is to obtain residue-specific characterization of the receptor binding sites and antigenic signatures on the envelope proteins of Venezuelan Equine Encephalitis virus (VEEV) and eventually for all alphaviruses. The Center for Disease Control and Prevention has classified VEEV as a bioterrorism agent, as it can be grown to high titers, and disseminated by aerosol. We will test the hypothesis that a comprehensive bioinformatics approach, combined with assays for cell binding, infection, and neutralization with monoclonal antibodies, will identify areas of the glycoproteins that determine antigenicity, fusion, and receptor interactions with host cells. We will establish a 3D-database of interacting regions of known complexes and new tools to locate areas with similar physical chemical properties in other proteins. The resulting trained neural networks or support vector machines will be used to identify areas of the envelope glycoproteins E1 and E2 of VEEV that are most likely to interact. As the 3D structures of E1 and E2 of VEEV have not been experimentally determined, we will model them on existing templates. We will map mutations of El and E2 that correlate with virulence on the models and analyze the structures of the surrounding areas. Mutants in selected areas will be assayed for their role in viral entry and fusion, using murine leukemia virus (MLV) pseudotypes that express the VEEV envelope proteins. This novel expression system permits study of the interacting sites of VEEV without the risks of dealing with infectious virus. Once the interacting areas of VEEV have been determined, our computational analysis will be extended to other pathogenic members of the alphavirus family. The resulting models can be used to design peptides mimicking the receptor recognition motifs, that may be the basis for the design of vaccines or inhibitors against alphaviruses.

描述（由申请人提供）：该项目的目的是获取委内瑞拉马脑炎病毒（VEEV）的包络蛋白上的受体结合位点和抗原特征的残留特异性表征，并最终用于所有α病毒。疾病控制与预防中心已将VEEV归类为生物恐怖剂，因为它可以生长到高滴度，并被气溶胶传播。我们将检验以下假设：一种综合的生物信息学方法，再加上与单克隆抗体中和细胞结合，感染和中和的测定，将确定确定确定抗原性，融合和受体相互作用的糖蛋白的区域。我们将建立一个已知复合物的相互作用区域和新工具的3D数据库，以定位其他蛋白质中具有相似物理化学特性的区域。由此产生的训练有素的神经网络或支持向量机将用于识别VEEV的包络糖蛋白E1和E2的区域，这些区域最有可能相互作用。由于尚未确定VEEV的E1和E2的3D结构，因此我们将在现有模板上对其进行建模。我们将绘制EL和E2的突变，这些突变与模型上的毒力相关并分析周围区域的结构。使用表达VEEV包膜蛋白的鼠白血病病毒（MLV）假型，将分析选定区域中的突变体在病毒进入和融合中的作用。这个新颖的表达系统允许研究VEEV的相互作用位点，而无需处理传染病。一旦确定了VEEV的相互作用领域，我们的计算分析将扩展到α病毒家族的其他致病成员。所得模型可用于设计模仿受体识别基序的肽，这可能是设计疫苗或抑制剂针对α病毒的基础。