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）：该项目将开始检查体育症病毒糖蛋白复合物的结构变化和功能域。融合孔的形成机理对于所有包膜病毒都是必需的。经典I类融合蛋白的详细检查已经阐明了流感，HIV和帕托马病毒的融合机制。 2007年将体育病毒糖蛋白作为I类融合蛋白的2007年分类所证明的，有关这些融合蛋白的结构，构象变化和生化表征的详细信息受到限制。竞技病毒家族被认为是一种新兴病毒，因为尽管该病毒在啮齿动物种群中是地方性的，但不断发现新菌株以感染人类。一些菌株，包括Lassa和Junin，引起毁灭性的出血发烧，发病率高和死亡率。此外，新的证据表明，美国和世界其他地区特有的淋巴细胞绒毛膜炎病毒可能是临床意义的疾病。目前没有有效的治疗剂或疫苗来治疗感染，仅LASSA发烧就会影响50万人，并造成西非5000人死亡。了解进入的机制，专门识别糖蛋白复合物（GPC）内的结构/功能域（GPC）将为未来的治疗提供新的靶标，并提供对研究研究的I类融合蛋白的基本理解。为此，该提案具有两个具体的目的：1）确定GP1亚基中的受体结合位点； 2）定义原位Lassa热糖蛋白复合物的几何形状。利用生化分析以及细胞对细胞融合测定，天然电子样品和最先进的冷冻电子层析成像，这是糖蛋白复合物所需的构象变化的模型，以诱导融合融合。未来使用基于结构设计的研究将能够靶向GPC中的特定构象，以产生有效的治疗剂和疫苗。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结合起来，以阐明其自然状态中体育症病毒融合的机制。这种合并的专业知识将为我在分子病毒学，计算生物学，生物化学和结构分析的交集的我未来的独立研究职业提供扎实的基础。通过协作，跨学科的方法实现我的项目目标，将成功地为我的未来研究团队领导和多样化的专业知识做好准备，这将被要求解决未来越来越复杂和多方面的生物学挑战。