A structural and biophysical study of the matrix proteins in influenza A/B viruses: Mechanisms of proton conduction and roles of protein-protein interactions

甲型/乙型流感病毒基质蛋白的结构和生物物理学研究：质子传导机制和蛋白质-蛋白质相互作用的作用

• 批准号: 9767794
• 负责人: Huong Tran Kratochvil
• 金额: $ 6.16万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9767794
• 关键词: Address; Advanced Development; Amantadine; Antigens; Binding; Biophysics; C-terminal; Capsid; Cells; Complex; Coupled; Data; Dependence; Development; Diffuse; Diffusion; Electrophysiology (science); Endosomes; Golgi Apparatus; Hemagglutinin; Human; Hydrogen Bonding; Immunoprecipitation; Influenza; Influenza A virus; Influenza B Virus; Investigation; Kinetics; Length; Life Cycle Stages; Link; M2 protein; Measures; Mediating; Membrane; Membrane Proteins; Methods; Modeling; Molecular Conformation; Motion; Movement; N-terminal; Nature; Pathway interactions; Peptides; Pharmaceutical Preparations; Play; Process; Proteins; Protons; Pump; Research; Resistance; Resolution; Roentgen Rays; Role; Specificity; Spectrum Analysis; Structural Protein; Structure; System; Time; Transmembrane Domain; Viral; Virus; Virus Replication; Water; Work; X-Ray Crystallography; anti-influenza; base; biophysical analysis; channel blockers; comparative; design; hydroxyl group; influenzavirus; insight; particle; premature; prevent; protein function; protein protein interaction; protein structure; single molecule; stem; stoichiometry; stop flow technique; targeted treatment; transmission process; viral RNA

Project Summary/Abstract The matrix protein M2 is a 97-residue homotetrameric protein that performs many important functions for the influenza virus. The primary function of this protein is to conduct protons to acidify the interior of the virus for the release of viral RNA from the capsid for replication and the transmission of infectious particles (11- 14). The transmembrane domain constitutes the functional core of the M2 protein, essential for the tetramerization and proton conductance (15), while the N- and C-terminal domains are necessary for viral budding (13, 14, 16, 17). From a biophysical perspective, M2 is also an interesting system to probe proton diffusion and pH-dependent conformational change as many of these structural and functional studies would yield insight into the proton conduction pathways of more complex proton channels and pumps. Because this protein is crucial for the viral life cycle, a detailed understanding of the M2 protein structure and its mechanisms of proton conduction will not only address the very fundamental processes of proton conduction and stabilization in membrane proteins, but will also aid in the development of targeted M2 channel blockers. The proposed research strategy herein seeks to elucidate the proton conduction mechanism in both AM2 and BM2 as well as probe its interactions with another matrix protein M1 to better understand the roles of these proteins in the replication and transmission of infectious influenza viruses. In this research, we propose to: 1. Link the proton-coupled conformational changes of AM2 to the kinetics of proton conduction and elucidate the hydrogen-bonding network of pore-lining residues and waters that stabilize the proton as it diffuses to the His tetrad to obtain a detailed picture of the conduction pathway through this protein. 2. Confirm the specificity and stoichiometry of binding of full-length AM2 to its interaction partner M1 and obtain a high-resolution structure of the resulting complex. 3. Determine the high-resolution structure of the functional core and of full-length BM2 to establish the conduction pathway through the protein and carry out a comparative analysis of the structures and mechanisms of proton conduction of AM2 and BM2.

项目总结/文摘