Solid-State NMR of Viral Fusion Peptides and Proteins

病毒融合肽和蛋白质的固态核磁共振

• 批准号: 7048792
• 负责人: David P Weliky
• 金额: $ 25.17万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7048792
• 关键词: HIV envelope protein gp41; Orthomyxoviridae; acidity /alkalinity; cell fusion; conformation; crosslink; host organism interaction; human immunodeficiency virus; intermolecular interaction; membrane activity; membrane fusion; membrane model; microorganism hemagglutinin; nuclear magnetic resonance spectroscopy; peptide structure; protein protein interaction; radiotracer; structural biology; virus infection mechanism; virus protein

DESCRIPTION (provided by applicant): The long-term goal of the proposed research is to understand with atomic-resolution detail the basis for viral fusion protein-induced membrane fusion. Fusion of viral and target cell membranes is a key step in infection for many viruses important in disease and a detailed understanding of fusion mechanisms should aid development of anti-viral therapeutics whose mode of action is fusion inhibition. The proposed research focuses on the gp41 and the hemagglutinin HA2 fusion proteins of the human immunodeficiency virus (HIV) and the influenza virus (IFV), respectively, because of the significance of the diseases caused by these viruses and because these proteins serve as prototypes for other class I viral fusion proteins. There is particular emphasis on the "fusion peptide" (FP) domains of the fusion proteins, which represent approximately 20-residue apolar regions at the N-termini of the proteins. Numerous biochemical and biophysical studies have shown that the FP plays a key role in fusion and that chemically synthesized peptides with the FP sequence can serve as useful model systems to understand some aspects of viral/target cell fusion. The long-term objectives will be achieved with four specific aims. Two of the aims focus on solid-state nuclear magnetic resonance (SSNMR) structural analysis of chemically cross-linked dimeric and trimeric HIV FPs in membranes and additional liquid-state NMR studies of these peptides in detergent micelles. The oligomeric topology of these C-terminal cross-linked peptides reflects the expected topology of FPs in the HIV gp41 fusion protein. The third aim focuses on: (a) determination of the insertion orientation of FPs in membranes using samples in which the membranes are oriented between stacked glass plates; and (b) probing FP insertion depths using SSNMR distance measurements between 13C nuclei in the FP and 31P and 19F nuclei in the lipid. The fourth specific aim focuses on SSNMR structural and insertion depth measurements for large HIV and IFV fusion protein domains which contain FPs.

描述（由申请人提供）：拟议研究的长期目标是用原子分辨率细节了解病毒融合蛋白诱导的膜融合的基础。病毒和靶细胞膜的融合是许多在疾病中重要的病毒感染的关键步骤，对融合机制的详细理解应有助于开发抗病毒疗法，其作用方式是融合抑制。拟议的研究重点是人类免疫缺陷病毒（HIV）（HIV）和流感病毒（IFV）的GP41和血凝素HA2融合蛋白，因为这些病毒引起的疾病的重要性以及这些蛋白质引起的疾病的意义，因为这些蛋白质是其他I型I型I型I病毒病毒型蛋白。特别强调了融合蛋白的“融合肽”（FP）结构域，该结构蛋白代表了蛋白质N末端的大约20个残留的极性区域。大量的生化和生物物理研究表明，FP在融合中起关键作用，并且具有FP序列的化学合成肽可以用作了解病毒/靶细胞融合的某些方面的有用模型系统。长期目标将以四个特定的目标实现。其中两个目的集中在膜中化学交联的二聚体和三聚体HIV FPS的固态核磁共振（SSNMR）结构分析以及这些肽在洗涤剂胶束中的其他液态液态NMR研究。这些C末端交联的肽的寡聚拓扑反映了HIV GP41融合蛋白中FPS的预期拓扑。第三目的重点是：（a）使用样品确定FPS在膜中插入方向的插入方向，其中将膜在堆叠的玻璃板之间定向； （b）使用脂质中FP和31p和19f核之间的SSNMR距离测量值探测FP插入深度。第四个特定目的侧重于大型HIV和IFV融合蛋白结构域的SSNMR结构和插入深度测量。