This application is for acquisition of a state-of-the-art nano-LC-MS/MS ion trap mass spectrometer, an amaZon ETD from Bruker Daltonics, which will provide accurate, reproducible and highly sensitive measurement of proteomics post-translational modifications. This instrument will bring significant enhancements to our Mass Spectrometry Core and will support the research efforts of multiple NIH-funded users. In addition to the high levels of sensitivity, the addition of electron transfer dissociation (ETD) asa fragmentation technique will facilitate novel proteomics discoveries in protein phosphorylation and acetylation as well as numerous other significant post-translational modifications (PTMs) such as protein adducts due to chronic oxidative stress and the resulting lipid peroxidation products 4-hydroxynonenal and acrolein. Multiple NIH-funded researchers at the University of Colorado Denver are proposed to share the use of this instrument. The primary justification for acquiring this instrument is to enhance these NIH-funded research projects in the area of proteomics discovery, utilizing the added sensitivity and ETD fragmentation as critical components. The investigators in this application possess the required experience and technological skill-set to maintain and properly use this instrument. Furthermore, the School of Pharmacy, where this instrument is proposed to be housed, is dedicated to supporting and developing its Mass Spectrometry Core and will provide the requisite service contracts for instrument maintenance and repair, as it does for each instrument housed within the Core facility. Furthermore, a cost recovery fee plan guarantees the financial viability for the instrumet over the long term. As detailed within this application, each major user demonstrates a rich and productive history in NIH-funded research. These NIH-funded projects range from neurodegenerative diseases to cancer and liver disease. A common denominator linking each of these projects is the need for highly sophisticated proteomics evaluation of PTMs and the impact these modifications play in altering basal physiology. The ability to examine these modifications with both collision induced dissociation (CID) and ETD fragmentation techniques will provide enhanced coverage of modified peptides. In particular, protein modifications which are labile under standard CID fragmentation conditions will be characterized with increased detail using ETD. These enhancements, coupled with an increase in sensitivity by 50-fold over our existing ion trap instrumentation will provide the capabilities for each shared user to reach both short and long-term research goals of studying altered proteomics under a wide variety of disease pathologies.
The instrument for this application is an amaZon ETD/CID ion trap mass spectrometer and nano liquid chromatography system from Bruker Daltonics which will provide accurate, reproducible and highly sensitive measurement of proteomics post-translational modifications. This instrument will support multiple NIH-funded research efforts in characterizing disease state-altered proteomics at the University of Colorado Denver Health Sciences Campus. Acquisition of this instrument would significantly enhance the efforts of the shared users in meeting both short and long-term research objectives in studying pathologies of neurodegenerative disease, liver disease and cancer.