Structures and lipid interactions of curvature-inducing membrane peptides by NMR

通过 NMR 观察曲率诱导膜肽的结构和脂质相互作用

• 批准号: 8418923
• 负责人: Mei Hong
• 金额: $ 28.21万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-01 至 2014-07-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8418923
• 关键词: Antibiotics; Antimicrobial Cationic Peptides; Antiviral Agents; Bacteria; Binding; Biological Process; C-terminal; Cardiolipins; Cell membrane; Cells; Cellular Membrane; Chimeric Proteins; Coiled-Coil Domain; Data; Dehydration; Development; Diffusion; Drug Delivery Systems; Excision; Family; Generations; Goals; HIV; HIV Envelope Protein gp41; Heterogeneity; Hydration status; Influenza Hemagglutinin; Integral Membrane Protein; Isotopes; Joints; Lead; Lipid Bilayers; Lipids; Magic; Measures; Membrane; Membrane Lipids; Membrane Proteins; Molecular Conformation; Nature; Paramyxovirus; Peptide Conformation; Peptides; Phase; Phosphatidylethanolamine; Phosphatidylglycerols; Proteins; Public Health; Recombinants; Resistance; Resolution; Roentgen Rays; Staging; Structure; Structure-Activity Relationship; Techniques; Tertiary Protein Structure; Transmembrane Domain; Viral; Viral Fusion Proteins; Virus; Virus Diseases; antimicrobial; antimicrobial peptide; base; design; insight; interest; microbial; mimetics; novel; novel vaccines; parainfluenza virus; peptide structure; physical property; protegrin PG-1; protein structure; public health relevance; research study; segregation; solid state nuclear magnetic resonance; tool; virus envelope

DESCRIPTION (provided by applicant): A diverse range of membrane peptides and proteins utilize membrane-curvature generation to carry out their biological function. Examples include cationic membrane peptides that disrupt microbial membranes or cross the membrane into cells by forming permanent or transient pores, and hydrophobic domains of viral fusion proteins that merge the virus envelope and the target cell membrane to cause viral entry. Elucidating how protein structures underlie membrane curvature generation has broad biomedical significance in enabling the design of more potent and resistance-free antibiotics and the development of new vaccines and antiviral drugs. The long-term objective of this project is to understand and quantify lipid-specific interactions of curvature-inducing membrane peptides. We will use solid-state NMR as our principal tool, because it is uniquely capable of simultaneously yielding high-resolution structures of membrane proteins and revealing the physical properties of the lipid membrane - curvature, dynamics, domain heterogeneity, and hydration - in which these peptides are embedded. We propose four specific aims. 1) We will investigate cationic-peptide- induced lipid clustering in bacteria-mimetic membranes. Potential segregation of anionic lipids from the main zwitterionic lipid of bacterial membranes, phosphatidylethanolamine, may be a key factor in promoting membrane curvature. Isotope-edited NMR experiments that probe lipid dynamics and peptide-lipid interactions will be conducted. Representative antimicrobial and cell-penetrating peptides such as PG-1 and HIV TAT will be examined. 2) We will develop 31P exchange NMR techniques to measure the curvature of mixed lipid membranes and the localization of cationic peptides in curvature-distinct domains. 3) We will determine the membrane-bound conformation, dynamics, depth of insertion, and oligomeric structure of the fusion peptide and the transmembrane domain of the paramyxovirus, PIV5. Structure information on PIV5 fusion protein will provide new insights into the mechanism of action of the important class I viral fusion proteins. 4) We will investigate the lipid interactions of the PIV5 fusion peptide using a variety of 2H, 31P, and 1H NMR experiments. Membrane curvature, fusion peptide localization, and membrane hydration will be characterized using both oriented and unoriented membranes, with the goal of understanding how the fusion peptide modifies the membrane structure to cause fusion.

描述（由申请人提供）：多种膜肽和蛋白质利用膜曲率产生来实现其生物学功能。实例包括破坏微生物膜或通过形成永久或瞬时孔穿过膜进入细胞的阳离子膜肽，以及合并病毒包膜和靶细胞膜以引起病毒进入的病毒融合蛋白的疏水结构域。阐明蛋白质结构如何构成膜曲率产生的基础，在设计更有效和无耐药性的抗生素以及开发新疫苗和抗病毒药物方面具有广泛的生物医学意义。本项目的长期目标是了解和量化曲率诱导膜肽的脂质特异性相互作用。我们将使用固态NMR作为我们的主要工具，因为它是唯一能够同时产生高分辨率的膜蛋白结构，并揭示脂质膜的物理性质-曲率，动力学，结构域异质性和水合作用-这些肽嵌入其中。我们提出了四个具体目标。1)我们将研究阳离子肽诱导的拟细菌膜中的脂质聚集。阴离子脂质与细菌膜的主要两性离子脂质磷脂酰乙醇胺的潜在分离可能是促进膜弯曲的关键因素。将进行同位素编辑的NMR实验，探测脂质动力学和肽-脂质相互作用。将检查代表性的抗菌肽和细胞穿透肽，如PG-1和HIV达特。2)我们将开发31 P交换NMR技术来测量混合脂质膜的曲率和曲率不同域中阳离子肽的定位。3)我们将确定膜结合的构象，动力学，插入深度，和寡聚体结构的融合肽和副粘病毒，PIV 5的跨膜结构域。PIV 5融合蛋白的结构信息将为重要的I类病毒融合蛋白的作用机制提供新的见解。4)我们将使用各种2 H、31 P和1H NMR实验研究PIV 5融合肽的脂质相互作用。膜曲率，融合肽的定位，和膜水合作用将使用定向和非定向膜的特点，与理解的融合肽如何修改膜结构，导致融合的目标。