Innovative Native Ion Mobility Approaches for Transformational Measurements in Structural Biology
用于结构生物学转化测量的创新天然离子淌度方法
基本信息
- 批准号:10477459
- 负责人:
- 金额:$ 28.85万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2020
- 资助国家:美国
- 起止时间:2020-09-05 至 2024-08-31
- 项目状态:已结题
- 来源:
- 关键词:ATP HydrolysisATP-Binding Cassette TransportersAddressAll-Trans-RetinolAmyloidosisAreaBindingBiologicalBiological AssayBiological ProcessBiophysicsBreathingCatalysisCerebrospinal FluidChargeChemistryComplementComplexCouplingCryoelectron MicroscopyDataDevelopmentDissociationEscherichia coliEventFormulationFourier TransformGerm-Line MutationGoalsHomoIndividualInvestigationIonsKineticsLifeLigand BindingLigandsLipid BindingLipidsMass Spectrum AnalysisMeasurementMembrane ProteinsMembrane Transport ProteinsModernizationModificationMolecular ConformationMonitorMotionNoiseNucleotidesOutcomePeptide ConformationPeriodicityPharmaceutical PreparationsPhysicsPhysiologicalPlasmaPositioning AttributePost-Translational Protein ProcessingPrealbuminProtein DynamicsProteinsProteomicsRecombinantsResearchResearch PersonnelResolutionRoentgen RaysSeminalShapesSignal TransductionSiteStructureSurfaceTechniquesTechnologyThermodynamicsThyroxineTimeTubeX-Ray Crystallographybasebiophysical techniquescomparativeexperiencefrontierimprovedinnovationinsightinstrumentinstrumentationion mobilitymass analyzermass spectrometermembermonomernon-Nativenovelpreservationprotein complexprotein expressionprotein foldingprotein purificationprotein structureprotein structure functionprototyperapid growthself assemblysmall moleculestoichiometrystructural biologysuccesstechnique developmenttime use
项目摘要
Project Summary
Advancements in biophysical techniques, such as X-ray and cryoEM, have undoubtedly accelerated
determination of protein structure. However, it still remains challenging to capture snapshots of protein
folding intermediates, including non-native states, and breathing motions that protein assemblies
undergo to perform their biological function. Moreover, understanding how molecules, such as lipids,
modulate protein structure and function is of paramount biological importance. Over the past two
decades, mass spectrometry (MS) of intact protein complexes, often referred to as native MS, has
emerged as an indispensable biophysical technique whereby non-covalent interactions and protein
structure are preserved within the mass spectrometer. Native MS is a rapid and sensitive technique
that has already provided invaluable information on subunit stoichiometry and topology, allostery and
cooperativity for individual ligand binding events, including their binding thermodynamics. The coupling
with ion mobility (IM), a separation technique based on molecule charge and shape, further enhances
the capabilities of native MS where it has enabled collision cross section (CCS) measurements for large
protein complexes, identification of different conformations for peptides and stabilizing ligands using
collision induced unfolding, and insight in folded and denatured structure(s) of proteins. However, low-
resolution commercial IM-MS instrumentation has not changed since its introduction 12 years ago.
Herein, this proposal seeks to develop transformative native IM-MS technologies with high-resolution
IM and MS capabilities that can address modern questions in structural biology, such as conformational
dynamics, including those that may have remained “hidden”, within membrane transporters under
turnover conditions. In order to achieve these transformative goals, an interdisciplinary team of
researchers whose expertise spans the fields of protein biophysics, expression and purification of
proteins inclusive of membrane proteins, as well as traditional protein structure characterization, such
as X-ray crystallography, has been assembled. Team members also possess decades of experience
in the field of mass spectrometry inclusive of fundamental ion chemistry/physics, seminal contributions
that have spawned MS proteomics, and related areas of analytical mass spectrometry and ion mobility-
mass spectrometry. Collectively, the background and expertise of this research team is uniquely
positioned to transform the field of IM-MS in the area of structural biology. In short, the proposed
transformative research will lead to forefront IM-MS instrumentation that is poised to provide
unprecedented insights into the structure and assembly of protein complexes and push the field into
new frontiers of research.
项目摘要
X射线和冷冻等生物物理技术的进步无疑已加速
蛋白质结构的测定。但是,捕获蛋白质快照仍然是挑战
折叠中间体,包括非本地状态和蛋白质组装的呼吸运动
要执行其生物学功能。此外,了解分子(例如脂质)如何
调节蛋白质结构和功能至关重要。在过去的两个
数十年来,完整蛋白质复合物的质谱(MS)通常称为天然MS
作为必不可少的生物物理技术出现,非共价相互作用和蛋白质
结构保留在质谱仪内。天然MS是一种快速而敏感的技术
这已经提供了有关亚基化学计量和拓扑,变构和拓扑结构的宝贵信息
单个配体结合事件的合作性,包括其结合热力学。耦合
使用离子迁移率(IM),一种基于分子电荷和形状的分离技术,进一步增强
本地MS的功能在其中启用了大型碰撞横截面(CCS)测量
蛋白质复合物,鉴定宠物的不同考虑和使用稳定配体的考虑因素
碰撞引起的展开,并在蛋白质的折叠和变性结构中见解。但是,低 -
自12年前引入以来,分辨率商业IM-MS仪器就没有改变。
在此,该提案旨在开发具有高分辨率的变革性的本地IM-MS技术
IM和MS能力可以解决结构生物学中的现代问题,例如构象
动态,包括可能仍然“隐藏”的动态,在膜转运器中
营业额条件。为了实现这些变革性目标,一个跨学科的团队
研究人员的专业知识涵盖了蛋白质生物物理学领域,表达和纯化的领域
包括膜蛋白的蛋白质以及传统的蛋白质结构表征,
作为X射线晶体学,已经组装了。团队成员还拥有数十年的经验
在包括基本离子化学/物理学的质谱领域中
产生了MS蛋白质组学以及分析质谱和离子迁移率的相关领域 -
质谱法。总的来说,该研究团队的背景和专业知识是独特的
定位于改变结构生物学领域的IM-MS领域。简而言之
变革性研究将导致最前沿的IM-MS仪器,该仪器被中毒以提供
对蛋白质复合物结构和组装的前所未有的见解,并将田间推入
研究的新边界。
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 }}
Brian Clowers其他文献
Brian Clowers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('Brian Clowers', 18)}}的其他基金
Tractable Tandem Ion Mobility Technology using Structures for Lossless Ion Manipulations and Photodissociation
使用无损离子操作和光解离结构的易处理串联离子淌度技术
- 批准号:
10386669 - 财政年份:2021
- 资助金额:
$ 28.85万 - 项目类别:
Tractable Tandem Ion Mobility Technology using Structures for Lossless Ion Manipulations and Photodissociation
使用无损离子操作和光解离结构的易处理串联离子淌度技术
- 批准号:
10322113 - 财政年份:2021
- 资助金额:
$ 28.85万 - 项目类别:
Tractable Tandem Ion Mobility Technology using Structures for Lossless Ion Manipulations and Photodissociation
使用无损离子操作和光解离结构的易处理串联离子淌度技术
- 批准号:
10548229 - 财政年份:2021
- 资助金额:
$ 28.85万 - 项目类别:
Innovative Native Ion Mobility Approaches for Transformational Measurements in Structural Biology
用于结构生物学转化测量的创新天然离子淌度方法
- 批准号:
10689746 - 财政年份:2020
- 资助金额:
$ 28.85万 - 项目类别:
Innovative Native Ion Mobility Approaches for Transformational Measurements in Structural Biology
用于结构生物学转化测量的创新天然离子淌度方法
- 批准号:
10042584 - 财政年份:2020
- 资助金额:
$ 28.85万 - 项目类别:
Innovative Native Ion Mobility Approaches for Transformational Measurements in Structural Biology
用于结构生物学转化测量的创新天然离子淌度方法
- 批准号:
10252003 - 财政年份:2020
- 资助金额:
$ 28.85万 - 项目类别:
相似国自然基金
基于超声多模态评价技术探讨肝脏靶向递送ABCA1新策略在动脉粥样硬化防治中的应用
- 批准号:81871357
- 批准年份:2018
- 资助金额:57.0 万元
- 项目类别:面上项目
基于SIRT1-LXR通路的化合物E4023抗动脉粥样硬化的作用及机制研究
- 批准号:81703503
- 批准年份:2017
- 资助金额:20.0 万元
- 项目类别:青年科学基金项目
基于LXRα-SREBP1-ABCA1/G1信号通路的益气活血化痰方调脂抗动脉粥样硬化机制研究
- 批准号:81774088
- 批准年份:2017
- 资助金额:55.0 万元
- 项目类别:面上项目
肝脏X受体激动剂干预β淀粉样蛋白诱导的视网膜炎性反应的作用及机制
- 批准号:81670881
- 批准年份:2016
- 资助金额:51.0 万元
- 项目类别:面上项目
新型ABCA1上调剂E17241改善糖脂代谢紊乱的机制研究
- 批准号:81573482
- 批准年份:2015
- 资助金额:50.0 万元
- 项目类别:面上项目
相似海外基金
Inhibition or evasion of P-glycoprotein-mediated drug transport
抑制或逃避 P-糖蛋白介导的药物转运
- 批准号:
10568723 - 财政年份:2023
- 资助金额:
$ 28.85万 - 项目类别:
ABC transporter-mediated secretion of capsular polysaccharides
ABC 转运蛋白介导的荚膜多糖分泌
- 批准号:
10412117 - 财政年份:2021
- 资助金额:
$ 28.85万 - 项目类别:
ABC transporter-mediated secretion of capsular polysaccharides
ABC 转运蛋白介导的荚膜多糖分泌
- 批准号:
10287699 - 财政年份:2021
- 资助金额:
$ 28.85万 - 项目类别: