Fusion Lumos Orbitrap System for Quantitative Proteomics and Metabolomics Applications

项目摘要 该应用程序用于安装Orbitrap Lumos Fusion液相色谱质谱计系统 在斯卡格斯药学和药学学院(SSPPS)的质谱学设施内 在科罗拉多大学安舒茨医学院。这台仪器将被用来推进几个 专注于定量和基于发现的蛋白质组学以及蛋白质组学后分析的翻译项目 翻译修饰(PTM)，如乙酰化，直接应用于人类健康和疾病。这个 仪器还将用于需要高分辨率以改进鉴定的代谢组学项目 未知化合物。用户的项目目前正在较旧的Q-TOF仪器上进行 无法获得多重蛋白质组学实验所需的蛋白质组覆盖或分辨率的深度， PTM分析，或精细分子结构的阐明。因此，迫切需要这种仪器。 Orbitrap Lumos对于预期的应用具有几个关键功能，包括特殊的灵敏度， 超高分辨率、MSN功能、电子转移解离(ETD)、定量 功能和发布得很好的方法。大卫·罗斯博士将利用轨道诱导器Lumos揭示机制 通过使用ETD进行深入的、定量的蛋白质组学和PTM分析，了解胰岛素敏感性 有可能影响针对代谢综合征的治疗。Thomas Delong博士将使用ETD进行新的工作 发现杂交肽以检查某些疾病的免疫学起源，如糖尿病和阿尔茨海默氏症 疾病。Ancldoquy博士将要求该仪器实现深层蛋白质组、代谢组和脂组 覆盖细胞外小泡，以改善药物输送系统。布朗医生需要一个高度敏感的 以及特定的氧化还原蛋白质组学方法来理解肥大细胞激活的调节。最后， Reisdorph博士和其他人将使用该仪器来支持使用其 能够进行多阶段实验，MSN，高分辨率，准确的质量。我们的长期目标是 利用Orbitrap Lumos进一步开发我们在乙酰组学、PTM定量和代谢组学方面的计划。 由于我们的核心目前缺乏任何类型的Orbitrap技术，我们的蛋白质组学和代谢组学服务受到了打击 一个平台；需要尖端的仪器才能把我们教员的研究带到下一个水平。这个 SSPPS在过去5年中投入了50多万美元(不包括设备)用于改善 由SSPPS支持了近20年的质谱学设施。我们的目标是 继续为我们的研究人员提供共享的质谱学资源，以支持当前和拟议的 美国国立卫生研究院资助的生物医学研究。这个轨道诱捕器对于该设施作为切割设备的继续发挥作用至关重要- 对于边缘企业和几个潜在影响较大的研究项目的成功。

Project Summary This application is for an Orbitrap Lumos Fusion liquid chromatography mass spectrometer system to be housed within the Mass Spectrometry Facility at the Skaggs School of Pharmacy and Pharmaceutical Sciences (SSPPS) at University of Colorado, Anschutz Medical Campus. This instrument will be used to advance several translational projects that focus on quantitative and discovery-based proteomics and the analysis of post- translational modifications (PTMs), such as acetylation, with direct application to human health and disease. The instrument will also be used for metabolomics projects that require high resolving power to improve identification of unknown compounds. The Users’ projects are currently being conducted on older, Q-TOF instrumentation that cannot acquire the depth of proteome coverage or resolution required for multiplex proteomics experiments, PTM analysis, or fine molecular structure elucidation. Hence there is an urgent need for this instrumentation. The Orbitrap Lumos has several critical features for the intended applications including exceptional sensitivity, ultra high resolving power, MSn capabilities, electron transfer dissociation (ETD), versatility in quantitative capabilities, and well-published methods. Dr. David Ross will utilize the Orbitrap Lumos to reveal mechanisms underlying insulin sensitivity through in-depth, quantitative proteomics and PTM analysis using ETD, with the potential to impact therapies directed at metabolic syndrome. Dr. Thomas Delong will employ ETD for de novo hybrid peptide discovery to examine immunological origins of certain diseases, such as diabetes and Alzheimer’s disease. Dr. Anchordoquy will require the instrument to achieve deep proteome, metabolome, and lipidome coverage of extracellular vesicles towards improved drug delivery systems. Dr. Brown requires a highly sensitive and specific redox proteomics approach to understanding regulation of mast cell activation. Finally, the instrument will be used by Dr. Reisdorph and others to support the identification of small molecules using its capabilities in multi-stage experiments, MSn, high resolution, and accurate mass. Our long-term objective is to utilize the Orbitrap Lumos to further develop our programs in acetylomics, PTM quantitation, and metabolomics. As our Core currently lacks any type of Orbitrap technology, our proteomics and metabolomics services have hit a plateau; cutting edge instrumentation is required to take the research of our faculty to the next level. The SSPPS has committed over $500,000 (not including equipment) in the past 5 years towards the improvements of mass spectrometry facility, which has been supported by SSPPS for almost 20 years. Our objective is to continue to provide our investigators with a shared mass spectrometry resource to support current and proposed NIH-funded biomedical research. This Orbitrap is vital to the continued functioning of the facility as a cutting- edge enterprise and for the success of several potentially high impact research projects.