Enabling Technologies to Characterize Biomolecules and Fluorophores in Controlled Micro-environments using Mass Spectrometry

使用质谱技术来表征受控微环境中的生物分子和荧光团

• 批准号: RGPIN-2020-05828
• 负责人: Jockusch, Rebecca
• 金额: $ 2.62万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=748352
• 关键词: Enabling; Technologies; Characterize; Biomolecules; Fluorophores

The Jockusch laboratory aims to create transformative tools and technologies that will have long-term impact on fields ranging from chemistry and physics to biology and medicine. This research program will advance understanding of how key molecules, such as proteins and DNA, are affected by their molecular environment. We primarily focus on building enabling technologies for mass spectrometry (MS), many of which exploit laser spectroscopy. Our work addresses critical challenges at heart of chemistry, biology and medicine. For example, proteins that are `misfolded' have costly impacts on health, due to their association with devastating diseases including Alzheimer's, Parkinson's and type II diabetes. Our contribution to combatting such protein-misfolding diseases is to develop and apply techniques that illuminate how different interactions contribute to protein folding, misfolding and aggregation. This will uncover interactions important for the development of efficacious drugs. In a related endeavor, we seek to improve the understanding of popular molecular sensors whose fluorescence is used to characterize key molecular states, including disease-correlated states. For example, we target understanding whether the best-known sensors for amyloid fibrils, which are late-stage misfolded protein aggregates, can report on the earlier-stage oligomers that are important therapeutic targets in Alzheimer's research. In order to dissect the intricate web of molecular interactions present in Nature, we begin by examining simplified systems whose complexity is then gradually built up in a carefully controlled fashion. We establish baseline behavior by characterizing key properties of bare, mass-selected ions. Then, pieces of the molecular environment (such as water molecules, metal ions, drugs or other binding partners) are added to form complexes and clusters, thus enabling discovery of how additional specific interactions modulate molecular properties of interest. Over the next five years, we aim to expand the suite of tools that can usefully be employed for molecular characterization. We primarily concentrate on developing new dimensions of readily-interpretable information from tandem MS analyses. For example, we made a flexible interface for laser spectroscopic experiments that enables the use of fluorescence and Förster resonance energy transfer (FRET) techniques to probe conformation, and other functional characteristics, of mass-selected ions. We also develop orthogonal probes of molecular characterization, such as gas-phase hydrogen-deuterium exchange. By combining information from these complementary techniques, a better picture will emerge of how molecular properties change with the local molecular environment. Overall, the techniques and tools developed in the Jockusch lab will be used to push scientific boundaries. This cutting-edge research program also provides an excellent training environment for the next generation of Canadian Scientists.

Jockusch实验室旨在创造革命性的工具和技术，这些工具和技术将对化学、物理、生物和医学等领域产生长期影响。这一研究项目将促进对关键分子（如蛋白质和DNA）如何受到其分子环境影响的理解。我们主要专注于为质谱（MS）建立使能技术，其中许多利用激光光谱学。我们的工作解决了化学、生物学和医学核心的关键挑战。例如，“错误折叠”的蛋白质会对健康造成代价高昂的影响，因为它们与阿尔茨海默病、帕金森病和II型糖尿病等毁灭性疾病有关。我们对对抗这种蛋白质错误折叠疾病的贡献是开发和应用技术，阐明不同的相互作用如何导致蛋白质折叠，错误折叠和聚集。这将揭示对开发有效药物至关重要的相互作用。在相关的努力中，我们寻求提高对流行分子传感器的理解，其荧光用于表征关键分子状态，包括疾病相关状态。例如，我们的目标是了解最著名的淀粉样原纤维传感器，它是晚期错误折叠的蛋白质聚集体，是否可以报告早期低聚物，这是阿尔茨海默病研究中重要的治疗靶点。为了剖析自然界中存在的分子相互作用的错综复杂的网络，我们首先检查简化的系统，然后以一种精心控制的方式逐渐建立其复杂性。我们通过描述裸的、大量选择的离子的关键特性来建立基线行为。然后，加入分子环境的片段（如水分子、金属离子、药物或其他结合伙伴）形成复合物和簇，从而能够发现额外的特定相互作用如何调节感兴趣的分子特性。在接下来的五年里，我们的目标是扩大一套工具，可以有效地用于分子表征。我们主要集中于从串联质谱分析中开发易于解释的信息的新维度。例如，我们为激光光谱实验制作了一个灵活的界面，可以使用荧光和Förster共振能量转移（FRET）技术来探测大量选择离子的构象和其他功能特征。我们还开发了分子表征的正交探针，如气相氢-氘交换。通过结合这些互补技术的信息，就可以更好地了解分子特性是如何随着局部分子环境的变化而变化的。总的来说，Jockusch实验室开发的技术和工具将用于推动科学界限。这个尖端的研究项目也为下一代加拿大科学家提供了良好的培训环境。