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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 M2是带有单个跨膜（TM）alpha螺旋拉伸的97个残基蛋白。 M2同二方在膜环境中形成质子选择通道。跨膜截面（M2TM）是序列SSDPLVVAASIIGIGILHLILWILDRL的25个氨基酸肽。流感病毒M2积分膜蛋白的离子通道活性在病毒生命周期中起重要作用。已经提出，四个跨膜螺旋中的每一个中的TRP 41对于M2的pH门控质子通道至关重要。发现在跨膜序列中其（H）的突变导致TRP（W）氨基酸的质子选择性和突变的丧失允许质子流出。因此，该项目的目的是将荧光相关光谱与激发态电子转移（TRP和标记的染料探针之间）结合使用，以探测吲哚环的构象动力学作为pH的函数。因此获得的信息应该使我们能够更好地了解M2通道的质子门控机制。我们还想研究发现可以阻断离子通道的小分子和rimantadine的作用。