Combined Mass Spectrometric and Optical Spectroscopic Investigation of Biological Molecules in Controlled Micro-environments

受控微环境中生物分子的质谱和光学光谱联合研究

• 批准号: RGPIN-2015-04599
• 负责人: Jockusch, Rebecca
• 金额: $ 3.28万
• 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: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=688884
• 关键词: Combined; Mass; Spectrometric; Optical; Spectroscopic

The development and application of new tools and techniques to identify molecules and study their properties can be transformative, with long-term impact on fields ranging from chemistry and physics to medicine and biology. The development of such tools is the aim of research in the Jockusch laboratory. In particular, we focus on the development of new techniques that extend the uses of mass spectrometry (MS) in order to enhance the understanding of how biological molecules such as proteins are affected by their environment. This is important because proteins play a central role in biology. For example, proteins that are `misfolded' have negative impacts on health, and are associated with many devastating diseases including Alzheimer's and Parkinson's diseases. However, proteins and other biomolecules do not generally act in isolation; their behavior is modulated by a complex web of molecular interactions. In order to dissect the complexity present in Nature, it is valuable to study simplified systems whose complexity can gradually be built up in a carefully controlled fashion. Thus, the Jockusch laboratory's approach to understanding the properties of biomolecules is to study them in a highly-controlled environment: isolated inside a mass spectrometer. This enables the separation of intrinsic molecular properties from effects of the molecular environment. In subsequent investigations, pieces of the molecular environment such as water molecules, metal ion, or other binding partners will be added to form complexes and clusters, thus enabling discovery of how additional specific interactions modulate or control the molecular properties of interest.***The Jockusch laboratory has added a new dimension of information to tandem mass spectrometry analysis by interfacing MS with a flexible interface for gas-phase laser spectroscopic experiments. In particular, we are spearheading the use of fluorescence and fluorescence resonance energy transfer (FRET) techniques to probe conformation and other functional characteristics of gaseous biomolecular ions and their complexes. FRET is an example of a transformative technique, developed by physicists and subsequently adopted by biologists for the study of protein conformation and dynamics. Its use enabled the study of even single molecules in solution - an amazing advance from ensemble techniques commonly used. The development of gas-phase FRET promises to be equally transformative. Only two other laboratories world-wide have capabilities and expertise similar to the Jockusch lab (at Harvard, and ETH Zurich), having implemented the powerful combination of mass spectrometry and fluorescence techniques. Not only will the techniques and tools developed in the Jockusch lab push the boundaries of MS-enabled science, this cutting-edge research program provides an excellent training environment for the next generation of Canadian Scientists.******

识别分子并研究其性质的新工具和新技术的开发和应用可能具有变革性，对化学、物理、医学和生物学等领域产生长期影响。开发这样的工具是Jockusch实验室的研究目标。特别是，我们专注于扩展质谱（MS）使用的新技术的开发，以增强对生物分子（如蛋白质）如何受其环境影响的理解。这一点很重要，因为蛋白质在生物学中起着核心作用。例如，“错误折叠”的蛋白质会对健康产生负面影响，并与许多毁灭性疾病有关，包括阿尔茨海默病和帕金森病。然而，蛋白质和其他生物分子通常不是孤立地起作用的；它们的行为受到分子相互作用的复杂网络的调节。为了剖析自然界中存在的复杂性，研究能够以一种谨慎控制的方式逐步建立复杂性的简化系统是有价值的。因此，Jockusch实验室了解生物分子特性的方法是在高度控制的环境中进行研究：在质谱仪中进行隔离。这使得从分子环境的影响分离固有的分子性质。在随后的研究中，分子环境的片段，如水分子、金属离子或其他结合伙伴将被添加到形成络合物和簇，从而能够发现额外的特定相互作用如何调节或控制感兴趣的分子性质。*** Jockusch实验室通过将质谱和气相激光光谱实验的柔性接口连接起来，为串联质谱分析增加了一个新的信息维度。特别是，我们率先使用荧光和荧光共振能量转移（FRET）技术来探测气态生物分子离子及其复合物的构象和其他功能特征。FRET是一种转化技术的例子，由物理学家开发，随后被生物学家用于蛋白质构象和动力学的研究。它的使用使得甚至可以研究溶液中的单个分子——这是通常使用的集成技术的一个惊人进步。气相FRET的发展承诺同样具有变革性。世界上只有另外两个实验室（哈佛大学和苏黎世联邦理工学院）拥有与Jockusch实验室类似的能力和专业知识，并实现了质谱法和荧光技术的强大结合。Jockusch实验室开发的技术和工具不仅将推动ms科学的边界，这个尖端的研究项目为下一代加拿大科学家提供了一个极好的培训环境。******