Elucidating the structural heterogeneity of differentially modified protein systems by tandem-ion mobility spectrometry / mass spectrometry methods.

通过串联离子迁移谱/质谱方法阐明差异修饰蛋白质系统的结构异质性。

• 批准号: 10455476
• 负责人: Christian Bleiholder
• 金额: $ 33.71万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10455476
• 关键词: 3-Dimensional; AIDS/HIV problem; Adopted; Amino Acid Sequence; Area; Biological; Communities; Data; Disease; Escape Mutant; Exhibits; Goals; HIV; Health; Heterogeneity; Human; Ions; Joints; Knowledge; Lead; Link; Mass Spectrum Analysis; Measurement; Measures; Methods; Mission; Molecular; Molecular Conformation; Motion; Outcome; Post-Translational Protein Processing; Preventive; Protein Sequence Analysis; Proteins; Public Health; Relaxation; Research; Resolution; Sequence Analysis; Spectrometry; Structure; System; Testing; Therapeutic; United States National Institutes of Health; Variant; Work; analytical method; combat; human disease; innovation; ion mobility; mutant; novel strategies; protein complex; protein function; protein protein interaction; protein structure; targeted treatment; three dimensional structure; two-dimensional; vaccine development

The precise mechanisms through which post-translational modifications modulate protein function are not clear nor is it understood how they are implicated in human diseases at the molecular level. The objective of this proposal is to develop a method that reveals how variations in sequence and post-translational modifications modulate the structural heterogeneity of proteins. Guided by our strong preliminary data, we will obtain the objective of this proposal by pursuing the following specific aims: 1) To enable structure-elucidation of differentially modified proteins by ion mobility spectrometry / mass spectrometry; and 2) To characterize the structural heterogeneity of differentially modified proteins by tandem-trapped ion mobility spectrometry / mass spectrometry. In the first Aim, we will develop a method that determines structures for differentially modified proteins and their assemblies, in particular for phosphorylated and glycosylated species. In the second Aim, we will develop an approach that reveals how protein structure depends on amino acid sequence and post- translational modifications. The research proposed in this application is innovative because it substantially advances from the status quo through unique computational and experimental methods that were recently developed in our lab, namely the Structure Relaxation Approximation and tandem-trapped ion mobility spectrometry/mass spectrometry methods. This contribution is significant because it is the first step towards a general analytical method that is expected to provide a molecular-level understanding of how changes in amino acid sequence and post-translational modifications are implicated in disease mechanisms. Ultimately, the results of the proposed work can be expected to significantly benefit a number of research areas relevant to the mission of the NIH, including the development of a vaccine against HIV/AIDS.

翻译后修饰调节蛋白质功能的确切机制尚不清楚 它也不了解它们如何与分子水平的人类疾病有关。这个目的 建议是开发一种方法，以揭示序列和翻译后修改的变化如何 调节蛋白质的结构异质性。在我们强大的初步数据的指导下，我们将获得 通过追求以下特定目的的目的：1） 通过离子迁移率 /质谱法差异修饰的蛋白质； 2）表征 通过串联离子迁移率 /质量的差异修饰蛋白的结构异质性 光谱法。在第一个目标中，我们将开发一种方法来确定差异修改的结构 蛋白质及其组件，特别是用于磷酸化和糖基化的物种。在第二个目标中，我们 将开发一种方法，该方法揭示蛋白质结构如何取决于氨基酸序列和后 翻译修改。本应用程序中提出的研究具有创新性，因为它基本上是 从现状到最近的独特计算和实验方法的进步 在我们的实验室中开发，即结构松弛近似和串联离子迁移率 光谱/质谱法。这项贡献很重要，因为它是迈向 一般的分析方法，预计将对氨基的变化提供分子级别的理解 酸序列和翻译后修饰与疾病机制有关。最终，结果 可以预期，拟议的工作将显着受益于许多与任务相关的研究领域 NIH，包括开发针对艾滋病毒/艾滋病的疫苗。

项目成果

Tandem Trapped Ion Mobility Spectrometry/Mass Spectrometry (tTIMS/MS) Reveals Sequence-Specific Determinants of Top-Down Protein Fragment Ion Cross Sections.

• DOI: 10.1021/acs.analchem.1c05171
• 发表时间: 2022-06-14
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Liu, Fanny C.;Kirk, Samuel R.;Caldwell, Kirsten A.;Pedrete, Thais;Meier, Florian;Bleiholder, Christian
• 通讯作者: Bleiholder, Christian

The role of solvation on the conformational landscape of α-synuclein

• DOI: 10.1039/d3an01680c
• 发表时间: 2023-11-17
• 期刊: ANALYST
• 影响因子: 4.2
• 作者: Kit，Melanie Cheung See;Cropley，Tyler C.;Webb，Ian K.
• 通讯作者: Webb，Ian K.

Perspective on the potential of tandem-ion mobility/mass spectrometry methods for structural proteomics applications

串联离子淌度/质谱方法在结构蛋白质组学应用中的潜力展望

• DOI: 10.3389/frans.2023.1106752
• 发表时间: 2023
• 期刊: Frontiers in Analytical Science
• 作者: Cropley, Tyler C.;Chai, Mengqi;Liu, Fanny C.;Bleiholder, Christian
• 通讯作者: Bleiholder, Christian