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: 7342783
• 负责人: David P Weliky
• 金额: $ 27.08万
• 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/7342783
• 关键词: Abbreviations; Acquired Immunodeficiency Syndrome; Affinity; Biochemical; Biological Assay; Biological Models; C-terminal; Catalysis; Cell Nucleus; Cell fusion; Cell membrane; Cells; Chemicals; Chimeric Proteins; Cholesterol; Circular Dichroism; Class; Coupled; Coupling; Data; Dependence; Depth; Detergents; Development; Disease; Electron Spin Resonance Spectroscopy; Environment; Erythrocytes; Ethers; Ethyl Ether; Glass; Glossary; Goals; HIV; Hemagglutinin; Human Parainfluenza Virus 1; Infection; Influenza; Intermediate Filament Proteins; Label; Link; Lipids; Liquid substance; Measurement; Membrane; Membrane Fusion; Membrane Proteins; Methods; Micelles; Molecular Conformation; Mutation; NMR Spectroscopy; Nuclear Magnetic Resonance; Peptides; Play; Point Mutation; Proteins; Rate; Research; Resolution; Role; Sampling; Structural Models; Structure; Techniques; Tertiary Protein Structure; Testing; Therapeutic; Time; Torsion; Vesicle; Viral; Viral Fusion Proteins; Viral Proteins; Virus; analog; analytical ultracentrifugation; base; crosslink; dimer; influenzavirus; magnetic field; monomer; novel; protein aminoacid sequence; prototype; research study; solid state

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 ~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. .

提出的研究的长期目标是了解原子分辨率细节的基础