Characterization of the arenavirus glycoprotein complex and mechanism of fusion

沙粒病毒糖蛋白复合物的表征和融合机制

基本信息

批准号：
9021590
负责人：
Melinda Ann Brindley
金额：
$ 10.8万
依托单位：
UNIVERSITY OF GEORGIA
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-03-01 至 2018-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9021590
关键词：
Address Affect Africa Alanine Antibodies Arenavirus Binding Sites Biochemical Biological Biological Assay Carbohydrates Categories Cell fusion Cells Cessation of life Chimeric Proteins Classification Comparative Study Complex Computational Biology Cysteine Data Development Disease Dystroglycan Early Intervention Electron Microscopy Electrophoresis Engineering Environment Event Exposure to Family Future GTPBP1 gene Geometry Glycoproteins Goals HIV Human In Situ Infection Influenza Junin virus Kinetics Lassa Fever Lassa fever virus Leadership Licensing Lymphocytic choriomeningitis virus Maps Measles Measles virus Mediating Modeling Molecular Conformation Molecular Target Molecular Virology Morbidity - disease rate Mutagenesis Mutation Old World Arenaviruses Paramyxovirus Pharmaceutical Preparations Population Process Reaction Research Rest Ribavirin Rodent Scanning Solid Staining method Stains Structural Biochemistry Structure Sulfides Surface Techniques Therapeutic Vaccines Viral Viral Hemorrhagic Fevers Virus Virus-like particle base career clinically significant comparative design effective therapy electron tomography flexibility geometric structure hemorrhagic fever virus innovation interdisciplinary approach mortality mutant novel therapeutics novel vaccines particle pathogen protein expression receptor binding research study therapeutic vaccine

项目摘要

DESCRIPTION (provided by applicant): This project will begin to examine the structural changes and the functional domains of the arenavirus glycoprotein complex. The mechanism of fusion pore formation is necessary for all enveloped viruses. Detailed examination of classical class-I fusion proteins have illuminated the fusion mechanisms of influenza, HIV and paramyxoviruses. As evidenced by the 2007 classification of the arenavirus glycoprotein as a class-I fusion protein, details about the structure, conformational changes, and biochemical characterization of these fusion proteins is limited. The arenavirus family in considered an emerging virus, because although the viruses are endemic in the rodent population, new strains continually are identified to infect humans. Some strains, including Lassa and Junin, cause devastating hemorrhagic fevers with high morbidity and mortality. In addition new evidence suggests that Lymphocytic Choriomeningitis virus, endemic to the US and the rest of the world may be responsible for clinically significant disease. There are no effective therapeutics or vaccines currently licensed to treat the infections, and Lassa fever alone affects half a million people each year and is responsible for 5000 deaths in West Africa. Understanding the mechanism of entry, specifically identifying the structural/functional domains within the glycoprotein complex (GPC) will provide new targets for future therapeutics, as well as provide a basic understanding of an understudied class-I fusion protein. Towards this end, this proposal has two Specific Aims: 1) Determine the receptor binding site in the GP1 subunit; 2) Define the geometry of the Lassa fever glycoprotein complex in situ. Utilizing biochemical analysis, as well as cell-to-cell fusion assays, native-electrophoresis, and state-of-the-art cryo-electron tomography, a model of the conformational changes required for the glycoprotein complex to induce fusion will emerge. Future studies using structure based designs will be able to target specific conformations in the GPC to produce effective therapeutics and vaccines. The project offers ample opportunity for me to utilize innovative electron microscopy techniques, and combine the structural data with advanced biochemical approaches to elucidate the mechanism of arenavirus fusion in their natural state. This combined expertise will provide a solid foundatio for my future independent research career at the intersection of molecular virology, computational biology, biochemistry and structural analysis. Reaching my project goals in a collaborative, interdisciplinary approach will successfully prepare me for future leadership of research teams and diverse expertise that will be required to solve increasingly complex and multifaceted biological challenges of the future.

描述（由申请人提供）：该项目将开始检查沙粒病毒糖蛋白复合物的结构变化和功能域。融合孔形成机制对于所有有包膜病毒都是必需的。对经典 I 类融合蛋白的详细检查阐明了流感、HIV 和副粘病毒的融合机制。正如 2007 年将沙粒病毒糖蛋白分类为 I 类融合蛋白所证明的那样，有关这些融合蛋白的结构、构象变化和生化特征的详细信息是有限的。沙粒病毒家族被认为是一种新兴病毒，因为尽管这些病毒在啮齿动物群体中流行，但不断发现新病毒株可感染人类。一些菌株，包括拉沙病毒和胡宁病毒，会引起毁灭性的出血热，发病率和死亡率很高。此外，新的证据表明，美国和世界其他地区流行的淋巴细胞性脉络丛脑膜炎病毒可能是导致临床上重大疾病的原因。目前尚无有效的疗法或疫苗获准治疗这些感染，仅拉沙热每年就影响 50 万人，并导致西非 5000 人死亡。了解进入机制，具体识别糖蛋白复合物 (GPC) 内的结构/功能域将为未来的治疗提供新的靶点，并提供对正在研究的 I 类融合蛋白的基本了解。为此，该提案有两个具体目标：1）确定GP1亚基中的受体结合位点； 2）原位定义拉沙热糖蛋白复合物的几何形状。利用生化分析以及细胞间融合测定、天然电泳和最先进的冷冻电子断层扫描，将出现糖蛋白复合物诱导融合所需的构象变化模型。未来使用基于结构的设计的研究将能够针对 GPC 中的特定构象来生产有效的治疗方法和疫苗。该项目为我提供了充分的机会，利用创新的电子显微镜技术，并将结构数据与先进的生化方法相结合，以阐明沙粒病毒在自然状态下的融合机制。这种综合的专业知识将为我未来在分子病毒学、计算生物学、生物化学和结构分析交叉领域的独立研究生涯奠定坚实的基础。通过协作、跨学科的方法实现我的项目目标将成功地为我未来的研究团队领导能力和解决未来日益复杂和多方面的生物挑战所需的多样化专业知识做好准备。