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）：该项目的目标是获得委内瑞拉马脑炎病毒（VEEV）以及最终所有甲病毒的包膜蛋白上的受体结合位点和抗原特征的残基特异性特征。美国疾病控制与预防中心已将 VEEV 归类为生物恐怖主义制剂，因为它可以生长到高滴度并通过气溶胶传播。我们将测试这样的假设：综合生物信息学方法，结合细胞结合、感染和单克隆抗体中和分析，将识别决定抗原性、融合以及受体与宿主细胞相互作用的糖蛋白区域。我们将建立一个已知复合物相互作用区域的 3D 数据库，并建立新工具来定位其他蛋白质中具有相似物理化学性质的区域。由此产生的经过训练的神经网络或支持向量机将用于识别 VEEV 的包膜糖蛋白 E1 和 E2 最有可能相互作用的区域。由于VEEV的E1和E2的3D结构尚未通过实验确定，我们将在现有模板上对其进行建模。我们将在模型上绘制与毒力相关的 El 和 E2 突变图谱，并分析周围区域的结构。将使用表达 VEEV 包膜蛋白的鼠白血病病毒 (MLV) 假型来分析选定区域的突变体在病毒进入和融合中的作用。这种新颖的表达系统允许研究 VEEV 的相互作用位点，而无需处理传染性病毒的风险。一旦确定了 VEEV 的相互作用区域，我们的计算分析将扩展到甲病毒家族的其他致病成员。由此产生的模型可用于设计模拟受体识别基序的肽，这可能是设计甲病毒疫苗或抑制剂的基础。