Solid-state NMR of influenza M2 protein in lipid bilayers

脂质双层中流感 M2 蛋白的固态 NMR

基本信息

  • 批准号:
    7939909
  • 负责人:
  • 金额:
    $ 27.75万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2009
  • 资助国家:
    美国
  • 起止时间:
    2009-09-30 至 2012-07-31
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): The M2 protein of influenza A virus forms a pH-gated proton channel that is important for viral infection and replication. The antiviral drug amantadine used to be effective in blocking this channel, until the recent emergence of a mutant, S31N, of the M2 transmembrane domain rendered the viruses completely resistant. The high- resolution structure of the M2 transmembrane peptide (M2TMP) is recently determined by X-ray crystallography and solution NMR. However, the two structures concluded dramatically different drug binding sites and noticeably different helix orientations and sidechain conformations. Since the X-ray and solution NMR structures were solved in detergents, these differences urge for high-resolution structural investigations in the more biologically relevant environment of lipid bilayers. Elucidation of the atomic-level structure of this important proton channel will help to develop new inhibitors to prevent future influenza pandemics. The broad, long-term objective of this work is to elucidate the structure and dynamics of the M2 protein in lipid bilayers in many of its functional states using solid- state NMR spectroscopy. We wish to understand how M2 conducts protons, how drug molecules block the channel, and how site-specific mutations alter the structure to evade drug binding. In the first funding period, we will focus on the transmembrane domain of the protein, and characterize the apo M2TMP in the closed state (high pH), the drug-complexed peptide in the closed state, and the apo peptide in the open state (low pH). We will also investigate the structure of the main amantadine-resistant mutant, S31N-M2TMP, and compare it with the structure of the wild-type peptide. Our main method is high-resolution magic-angle spinning (MAS) solid-state NMR, which allows atomic-resolution structural information to be obtained from unoriented hydrated lipid membrane samples. Based on our preliminary data, we hypothesize that a key element of M2TMP is its conformational flexibility, which is manifested as drug- and pH-induced backbone and sidechain conformational changes, mobility changes, and membrane-induced helix orientation changes. We will test this general hypothesis by measuring chemical shift perturbations, 13C and 15N linewidths, nuclear spin relaxation times, and the helix orientation in various states of the peptide. Experiments at physiological temperature will characterize the dynamic conformational fluctuations of the peptide, while low-temperature experiments will yield the average conformation and conformational distribution. We will further determine intermolecular distances between the drug and the peptide using both quantitative dipolar recoupling experiments and semi-quantitative spin diffusion techniques. Our goal is to obtain a high-resolution structure of bilayer-bound M2TMP with both backbone and sidechain constraints, and to elucidate the structural differences due to drug binding pH change, and mutation. PUBLIC HEALTH RELEVANCE: Atomic-resolution structure determination of the M2 protein of influenza A viruses in lipid bilayers is directly relevant to treating influenza infection and preventing future flu pandemics in the US and worldwide. The high-resolution structural information will be crucial for the design of new antiviral drugs to target the drug-resistant mutant proton channel, S31N-M2 in influenza A viruses.
描述(由申请人提供):甲型流感病毒M2蛋白形成ph门控质子通道,对病毒感染和复制很重要。抗病毒药物金刚烷胺过去可以有效阻断这一通道,直到最近出现的M2跨膜结构域的突变体S31N使病毒完全具有耐药性。最近用x射线晶体学和溶液核磁共振测定了M2跨膜肽(M2TMP)的高分辨率结构。然而,这两种结构的药物结合位点明显不同,螺旋取向和侧链构象也明显不同。由于在洗涤剂中解决了x射线和溶液核磁共振结构,这些差异迫切需要在更具有生物学相关性的脂质双分子层环境中进行高分辨率结构研究。阐明这一重要质子通道的原子水平结构将有助于开发新的抑制剂,以预防未来的流感大流行。这项工作的长期目标是利用固态核磁共振波谱技术阐明脂质双分子层中M2蛋白在许多功能状态下的结构和动力学。我们希望了解M2如何传导质子,药物分子如何阻断通道,以及位点特异性突变如何改变结构以逃避药物结合。在第一期资助期内,我们将重点研究该蛋白的跨膜结构域,对封闭状态(高pH)的载脂蛋白M2TMP、封闭状态的药物络合肽和开放状态(低pH)的载脂蛋白肽进行表征。我们还将研究主要的金刚烷胺抗性突变体S31N-M2TMP的结构,并将其与野生型肽的结构进行比较。我们的主要方法是高分辨率魔角旋转(MAS)固态核磁共振,它允许从无取向水合脂膜样品中获得原子分辨率的结构信息。根据我们的初步数据,我们假设M2TMP的一个关键因素是其构象灵活性,表现为药物和ph诱导的主链和侧链构象改变、迁移率改变和膜诱导的螺旋取向改变。我们将通过测量化学位移扰动、13C和15N线宽、核自旋弛豫时间和肽的各种状态下的螺旋取向来测试这一一般假设。生理温度下的实验将表征肽的动态构象波动,低温下的实验将得到平均构象和构象分布。我们将进一步利用定量偶极重偶联实验和半定量自旋扩散技术确定药物和肽之间的分子间距离。我们的目标是获得具有主链和侧链约束的双层结合M2TMP的高分辨率结构,并阐明由于药物结合pH变化和突变导致的结构差异。公共卫生相关性:甲型流感病毒脂质双层中M2蛋白的原子分辨率结构测定与治疗流感感染和预防未来在美国和世界范围内的流感大流行直接相关。高分辨率的结构信息对于设计针对甲型流感病毒中耐药突变质子通道S31N-M2的新型抗病毒药物至关重要。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

Mei Hong其他文献

Mei Hong的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('Mei Hong', 18)}}的其他基金

M. Hong RT&D
洪明 RT
  • 批准号:
    10224250
  • 财政年份:
    2019
  • 资助金额:
    $ 27.75万
  • 项目类别:
Tau structure and dynamics in Alzheimer's disease
阿尔茨海默病中的 Tau 结构和动力学
  • 批准号:
    10659553
  • 财政年份:
    2018
  • 资助金额:
    $ 27.75万
  • 项目类别:
Molecular structures of tau aggregates studied by solid-state NMR
通过固态核磁共振研究 tau 聚集体的分子结构
  • 批准号:
    10230898
  • 财政年份:
    2018
  • 资助金额:
    $ 27.75万
  • 项目类别:
Solid-state NMR of the influenza M2 protein in lipid bilayers
脂质双层中流感 M2 蛋白的固态 NMR
  • 批准号:
    8508272
  • 财政年份:
    2009
  • 资助金额:
    $ 27.75万
  • 项目类别:
Solid-state NMR of the influenza M2 protein in lipid bilayers
脂质双层中流感 M2 蛋白的固态 NMR
  • 批准号:
    9231933
  • 财政年份:
    2009
  • 资助金额:
    $ 27.75万
  • 项目类别:
Structures and Dynamics of Proton and Cation-Dependent Channels and Transporters
质子和阳离子依赖性通道和转运蛋白的结构和动力学
  • 批准号:
    10659039
  • 财政年份:
    2009
  • 资助金额:
    $ 27.75万
  • 项目类别:
Structures and Dynamics of Proton and Cation-Dependent Channels and Transporters
质子和阳离子依赖性通道和转运蛋白的结构和动力学
  • 批准号:
    10296879
  • 财政年份:
    2009
  • 资助金额:
    $ 27.75万
  • 项目类别:
Quadruple-resonance HFXY 1.3 mm CP-MAS probe for a solid-state NMR wide-bore magnet
用于固态 NMR 大口径磁体的四共振 HFXY 1.3 mm CP-MAS 探头
  • 批准号:
    10798817
  • 财政年份:
    2009
  • 资助金额:
    $ 27.75万
  • 项目类别:
Solid-state NMR of the influenza M2 protein in lipid bilayers
脂质双层中流感 M2 蛋白的固态 NMR
  • 批准号:
    9306548
  • 财政年份:
    2009
  • 资助金额:
    $ 27.75万
  • 项目类别:
Solid-state NMR of the influenza M2 protein in lipid bilayers
脂质双层中流感 M2 蛋白的固态 NMR
  • 批准号:
    8211383
  • 财政年份:
    2009
  • 资助金额:
    $ 27.75万
  • 项目类别:

相似海外基金

Attenuation of postoperative cognitive dysfunction by amantadine
金刚烷胺减轻术后认知功能障碍
  • 批准号:
    9125718
  • 财政年份:
    2015
  • 资助金额:
    $ 27.75万
  • 项目类别:
Assessing arousal regulation in post-stroke apathy using amantadine and EEG.
使用金刚烷胺和脑电图评估中风后冷漠的唤醒调节。
  • 批准号:
    8874363
  • 财政年份:
    2014
  • 资助金额:
    $ 27.75万
  • 项目类别:
Assessing arousal regulation in post-stroke apathy using amantadine and EEG.
使用金刚烷胺和脑电图评估中风后冷漠的唤醒调节。
  • 批准号:
    8678271
  • 财政年份:
    2014
  • 资助金额:
    $ 27.75万
  • 项目类别:
Assessing arousal regulation in post-stroke apathy using amantadine and EEG.
使用金刚烷胺和脑电图评估中风后冷漠的唤醒调节。
  • 批准号:
    9027865
  • 财政年份:
    2014
  • 资助金额:
    $ 27.75万
  • 项目类别:
Generation of a rabbit monoclonal antibody that discriminates between acetyl amantadine and amantadine
区分乙酰金刚烷胺和金刚烷胺的兔单克隆抗体的产生
  • 批准号:
    452021-2013
  • 财政年份:
    2013
  • 资助金额:
    $ 27.75万
  • 项目类别:
    Engage Grants Program
Amantadine + rTMS as a Neurotherapeutic for Disordered Consciousness after TBI
金刚烷胺 rTMS 作为 TBI 后意识障碍的神经治疗药物
  • 批准号:
    8712528
  • 财政年份:
    2013
  • 资助金额:
    $ 27.75万
  • 项目类别:
Amantadine + rTMS as a Neurotherapeutic for Disordered Consciousness after TBI
金刚烷胺 rTMS 作为 TBI 后意识障碍的神经治疗药物
  • 批准号:
    8584136
  • 财政年份:
    2013
  • 资助金额:
    $ 27.75万
  • 项目类别:
Évaluation des effets des psychostimulants amantadine et méthylphénidate sur la sortie comateuse et la récupération cognitive chez les patients ayant subi un traumatisme crânio-cérébral modéré à sévère
精神兴奋剂金刚烷胺和甲基苯酯对患者认知恢复和颅内创伤的影响评估
  • 批准号:
    252262
  • 财政年份:
    2012
  • 资助金额:
    $ 27.75万
  • 项目类别:
    Studentship Programs
AMANTADINE FOR PREVENTION OF WEIGHT GAIN IN FIRST EPIDSODE ADULT PSYCHOTIC SUBJ
金刚烷胺用于预防成人精神病首发患者体重增加
  • 批准号:
    7716804
  • 财政年份:
    2008
  • 资助金额:
    $ 27.75万
  • 项目类别:
Structural basis for influenza amantadine-resistance
流感金刚烷胺耐药的结构基础
  • 批准号:
    7458090
  • 财政年份:
    2006
  • 资助金额:
    $ 27.75万
  • 项目类别:
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了