Structural determinants of membrane fusion by HSV-1 gB

HSV-1 gB 膜融合的结构决定因素

• 批准号: 6962233
• 负责人: Ekaterina Heldwein
• 金额: $ 16.87万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6962233
• 关键词: Herpesviridae disease; antiviral agents; gene mutation; glycoprotein structure; herpes simplex virus 1; membrane fusion; oligonucleotides; phenotype; protein purification; protein structure function; virus envelope; virus infection mechanism; virus protein

DESCRIPTION (provided by applicant): Herpes simplex viruses infect their hosts for life, causing cold sores (HSV-1), eye and genital infections (HSV-1 and -2), and encephalitis. Furthermore, they can wreak havoc in immuno-compromised individuals or in newborns. Host cell entry by HSV-1 requires four viral envelope glycoproteins: gB, gD, and gH/gL complex. Binding of gD to a cell-surface receptor, which triggers fusion events, has been studied crystallographically. By contrast, no structural information is available on the other three proteins, and little is known about how they participate in cell entry. Knowing the structures of all the components of the membrane fusion machinery is a necessary step in unveiling the mechanism of HSV cell fusion. Our long-term goal is to elucidate the mechanism of Herpes virus entry into host cells at the atomic level and to use this knowledge to design effective inhibitors of viral entry into cells for preventative and therapeutic purposes. In preliminary studies, we have crystallized the ectodomain of gB and are currently determining its structure. In this application we propose biochemical and structural studies of additional domains and larger forms of gB, in order to build a more complete picture of its activities and interactions The three specific aims are 1) to determine the structure of the cytoplasmic domain of gB; 2) to produce pure, soluble forms of gB encompassing multiple domains for structural studies; and 3) to study the mechanism of the antiviral effect of a G-quartet oligonucleotide crystallographically. This structural information will provide the framework for elucidating the function of gB in cell entry. For example, known functional mutations will be mapped onto the structure. Moreover, strategies developed for producing soluble multidomain forms of gB for structural studies will allow us to approach gH/gL and, eventually, to assemble multi-protein complexes.

描述（由申请人提供）：单纯疱疹病毒感染其生命的宿主，引起唇疱疹（HSV-1），眼睛和生殖器感染（HSV-1和-2）和脑炎。此外，他们可能会对免疫受损的人或新生儿造成严重破坏。 HSV-1的宿主细胞进入需要四个病毒包膜糖蛋白：GB，GD和GH/GL复合物。 GD与触发融合事件的细胞表面受体的结合已在结晶上进行了研究。相比之下，其他三种蛋白质上没有任何结构信息，并且关于它们如何参与细胞进入的知之甚少。了解膜融合机械的所有组件的结构是揭示HSV细胞融合机制的必要步骤。我们的长期目标是阐明疱疹病毒在原子水平上进入宿主细胞的机制，并利用这些知识来设计有效的病毒进入细胞的抑制剂，以进行预防和治疗目的。在初步研究中，我们已经结晶了GB的外生域，目前正在确定其结构。在此应用中，我们提出了对其他域和更大形式的GB的生化和结构研究，以构建其活动和相互作用的更完整的图片，三个特定目标是1）确定GB的细胞质结构域的结构； 2）生产纯净的GB的可溶性形式，其中包含多个结构研究的多个领域； 3）研究G-Quartet寡核苷酸的抗病毒作用的机制。这些结构信息将为阐明GB在细胞进入中的功能提供框架。例如，已知的功能突变将映射到结构上。此外，为生产可溶性多域形式GB制定的策略将使我们能够接近GH/GL，并最终可以组装多蛋白复合物。