PROBING THE PROTON GATING DYNAMICS OF THE INFLUENZA VIRUS M2 CHANNEL USING FCS

使用 FCS 探测流感病毒 M2 通道的质子门动力学

• 批准号: 7955458
• 负责人: WILLIAM DEGRADO
• 金额: $ 0.96万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7955458
• 关键词: Amantadine; Amino Acids; Computer Retrieval of Information on Scientific Projects Database; Dyes; Electron Transport; Environment; Fluorescence; Funding; Goals; Grant; Indoles; Institution; Integral Membrane Protein; Ion Channel; Label; Laboratories; Life Cycle Stages; Membrane; Mutation; Optics; Play; Process; Proteins; Protons; Research; Research Personnel; Resources; Rimantadine; Role; Source; Spectrum Analysis; Stretching; United States National Institutes of Health; Virus; influenzavirus; protein aminoacid sequence; small molecule

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. M2 is a 97 residue protein with a single transmembrane (TM) alpha helical stretch. M2 homotetramerizes to form a proton selective channel in a membrane environment. The transmembrane section (M2tm) is a 25 amino acid peptide of the sequence SSDPLVVAASIIGILHLILWILDRL. The ion channel activity of the influenza virus M2 integral membrane protein plays an important role in the life cycle of the virus. It has been proposed that Trp 41 in each one of the four transmembrane helices is crucial for the pH-gated proton channel of M2. It was found that mutation of teh His (H) in the transmembrane sequence leads to a loss of the proton selectivity and mutation of the Trp (W) amino acid allows for the outflow of protons. Thus, the goal of this project is to use fluorescence correlation spectroscopy in conjunction with excited state electron transfer (between Trp and a labeled dye probe) to probe the conformational dynamics of the indole rings as a function of pH. The information thus obtained should allow us to gain a better understanding of the proton gating mechanism of the M2 channel. We also would like to investigate the action of the small molecules Amantadine and Rimantadine which were found to block the ion channel.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 M2 是一种具有单跨膜 (TM) α 螺旋延伸的 97 个残基蛋白质。 M2 同四聚化，在膜环境中形成质子选择性通道。跨膜部分 (M2tm) 是序列 SSDPLVVAASIIGILHLILWILDRL 的 25 个氨基酸肽。流感病毒M2整合膜蛋白的离子通道活性在病毒的生命周期中起着重要作用。有人提出，四个跨膜螺旋中每一个的 Trp 41 对于 M2 的 pH 门控质子通道至关重要。研究发现跨膜序列中His (H) 的突变会导致质子选择性丧失，而Trp (W) 氨基酸的突变会导致质子流出。因此，该项目的目标是使用荧光相关光谱与激发态电子转移（Trp 和标记染料探针之间）结合来探测吲哚环的构象动力学作为 pH 的函数。由此获得的信息应该能让我们更好地理解 M2 通道的质子门控机制。我们还想研究小分子金刚烷胺和金刚乙胺的作用，它们被发现可以阻断离子通道。