Fusion Lumos Orbitrap System for Quantitative Proteomics and Metabolomics Applications

项目总结

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.