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: 10224264
• 负责人: Christian Bleiholder
• 金额: $ 33.42万
• 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 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10224264
• 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 我们也不了解它们如何在分子水平上与人类疾病相关。此举的目的 建议开发一种方法来揭示序列和翻译后修饰的变化 调节蛋白质的结构异质性。在我们强有力的初步数据的指导下，我们将获得 通过追求以下具体目标来实现本提案的目标： 1) 实现结构阐明 通过离子迁移谱/质谱法对蛋白质进行差异修饰； 2) 表征 通过串联捕获离子迁移谱/质量分析差异修饰蛋白质的结构异质性 光谱测定法。在第一个目标中，我们将开发一种确定差异修饰结构的方法 蛋白质及其组装体，特别是磷酸化和糖基化物种。在第二个目标中，我们 将开发一种方法，揭示蛋白质结构如何取决于氨基酸序列和后处理 翻译修饰。本申请中提出的研究具有创新性，因为它实质上 通过最近出现的独特计算和实验方法从现状中取得进展 我们实验室开发的，即结构弛豫近似和串联捕获离子淌度 光谱分析/质谱分析方法。这一贡献意义重大，因为这是迈向 通用分析方法，有望提供分子水平上氨基如何变化的理解 酸序列和翻译后修饰与疾病机制有关。最终结果是 预计拟议工作将大大有益于与特派团相关的一些研究领域 NIH 的工作，包括开发艾滋病毒/艾滋病疫苗。