Enabling Top-Down Proteomics through Material Chemistry and Nanotechnology

通过材料化学和纳米技术实现自上而下的蛋白质组学

• 批准号: 9336949
• 负责人: Ying Ge
• 金额: $ 29.55万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-25 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9336949
• 关键词: Acids; Address; Alternative Splicing; Binding; Biological; Biological Markers; Categories; Cell physiology; Chemistry; Chromatography; Code; Collaborations; Complex; Computer software; DNA Sequence Alteration; Detection; Development; Digestion; Disadvantaged; Disease; Early Diagnosis; Face; Foundations; Genes; Genetic Variation; Human; Human Characteristics; Hydrophobic Interactions; Ligands; Liquid Chromatography; Marriage; Mass Spectrum Analysis; Membrane; Membrane Proteins; Methods; Molecular; Molecular Sieve Chromatography; Mutation; Nanotechnology; Peptides; Performance; Phosphoproteins; Phosphotransferases; Play; Post-Translational Protein Processing; Prevention; Proteins; Proteome; Proteomics; RNA Splicing; Research; Resolution; Role; Sampling; Signal Transduction; Silicon Dioxide; Solubility; Specificity; System; Technology; Variant; base; disease diagnosis; human disease; innovation; insight; interdisciplinary approach; nanomaterials; nanoparticle; new therapeutic target; novel; novel strategies; protein expression; success; surfactant; tool

Summary Proteomics is essential for deciphering how biomolecules interact as a system and for understanding the functions of cellular systems in human diseases. However, the unique characteristics of the human proteome, which include the large dynamic range of protein expression and the extreme complexity resulting from a plethora of post-translational modifications (PTMs) and sequence variations, make such analyses challenging. A comprehensive analysis of all proteoforms in the human proteome that arise from genetic variations, alternative splicing, and PTMs, is essential for gaining a transformative understanding of disease mechanisms and identifying new therapeutic targets. The emerging top-down mass spectrometry (MS)-based proteomics, which is based on analysis of intact proteins, is arguably the most powerful method to comprehensively characterize proteoforms to decipher the PTM codes together with genetic variations. However, top-down MS-based proteomics still faces significant challenges in terms of protein solubility, protein separation, and detection of low- abundance proteins. In this multiple-PI project, we will develop novel approaches enabled by nanotechnology and materials chemistry to overcome the challenges facing top-down proteomics in a comprehensive manner. The specific objectives of this proposal are: 1) To develop novel top-down MS-compatible surfactants that can effectively solubilize all categories of proteins including membrane proteins. 2) To develop novel chromatography materials based on mesoporous silica nanomaterials with uniform pore sizes and new strategies for high- resolution multi-dimensional liquid chromatography for effective separation of intact proteins. 3). To develop functionalized multivalent nanoparticles for highly specific enrichment of low-abundance proteins and proteins with PTMs. Our highly interdisciplinary approach is a perfect marriage between materials chemistry/nanotechnology and top-down MS-based proteomics. It is built on an existing collaboration between the two PIs that has led to significant preliminary results. We envision the development of the proposed technologies will significantly advance the burgeoning field of top-down proteomics. The success in our research will provide innovative tools for gaining transformative insights into the molecular basis of diseases and developing new strategies for the early diagnosis, prevention, and better treatment of human diseases.

总结 蛋白质组学对于破译生物分子作为一个系统如何相互作用以及理解生物分子的功能是必不可少的。 细胞系统在人类疾病中的功能。然而，人类蛋白质组的独特特征， 包括蛋白质表达的大动态范围和由过多的 翻译后修饰（PTM）和序列变异的复杂性使得这种分析具有挑战性。一 全面分析人类蛋白质组中由遗传变异引起的所有蛋白质型， 剪接和PTM对于获得对疾病机制的变革性理解至关重要， 寻找新的治疗靶点新兴的基于自上而下质谱（MS）的蛋白质组学， 基于对完整蛋白质的分析，可以说是全面表征 蛋白质形式来破译PTM代码以及遗传变异。然而，基于MS的自顶向下 蛋白质组学在蛋白质溶解度、蛋白质分离和低浓度蛋白质的检测方面仍然面临重大挑战。 蛋白质丰度在这个多PI项目中，我们将开发由纳米技术实现的新方法 和材料化学，以全面的方式克服自上而下的蛋白质组学所面临的挑战。 该建议的具体目标是：1）开发新型自上而下的MS相容性表面活性剂， 有效地溶解所有种类的蛋白质，包括膜蛋白。2)开发新型色谱 材料基于具有均匀孔径的介孔二氧化硅纳米材料，以及用于高性能纳米材料的新策略。 分辨率多维液相色谱法，用于完整蛋白质的有效分离。3）。发展 用于高特异性富集低丰度蛋白质和蛋白质的官能化多价纳米颗粒 用PTM。我们高度跨学科的方法是材料之间的完美结合 化学/纳米技术和自上而下的MS蛋白质组学。它是建立在现有的合作， 这两项PI已经取得了重要的初步成果。我们设想拟议的发展 技术将大大推进自上而下的蛋白质组学这一新兴领域。我们研究的成功 将提供创新工具，以获得对疾病分子基础的变革性见解， 为人类疾病的早期诊断、预防和更好的治疗制定新的战略。