This shared instrumentation proposal seeks funding for the purchase of an AB Sciex next generation TripleTOF 5600 quadrupole orthogonal time-of-flight mass spectrometer. This mass spectrometer will be part of an integrated nano-2DLC MS/MS system to be completed through independent funds already committed by the Buck Institute for this purpose. In brief, we propose to operate this integrated TripleTOF mass spectrometry system exclusively for the on-line separation and analysis of complex peptide and protein mixtures as part of large group of 12 independent principal investigators from the Buck Institute and 6 external investigators that represent a total of seven institutions or universities. Together, these researches bring together 21 NIH funded grants, all of which have significant needs for state-of-the-art mass spectrometry for various proteomics applications that require the highest sampling efficiency, resolution and sensitivity. The most critical features of this new instrumentation will be to the high resolution capabilities of data acquisition in both MS1 (40,000) and MS2 (15,000 - 35,000), the number of tandem mass spectra that can be acquired in a given time (typically 35 scans /sec), the exquisite sensitivity (low attomoles) and high dynamic range. These operating parameters will also allow several unique data acquisition scans, including SWATH and pseudo-MRM, and novel quantitation approaches that uses both MS1 and MS2 ion intensity data. The unique capabilities of this instrumentation will be directed at a diverse array of protein and proteomic application where mass spectrometry is used for identification, discovery, and global quantitation in very complex biological samples. The identification and quantitation of biologically important enzymatic and non-enzymatic posttranslational modifications will be a critical use of this new mass spectrometer, including protein phosphorylation, lysine-acetylation, and oxidative damage, among others. Overall, the acquisition of this instrument to the Chemistry &Mass Spectrometry Core at the Buck Institute for Research on Aging will greatly improve the throughput, experimental design, and discovery rate of over 19 projects that are examining key issues of human health and health span, including the biology of aging, neurodegenerative disease, diabetes, cancer, disease biomarker discovery, and infectious diseases.

Public Health Relevance

of this new instrumentation to human health is significant with over 19 independent researchers at seven institutions that will use this new technology to improve their ongoing studies in the basic biology of aging and age-related diseases, including Huntington, Alzheimer and Parkinson's disease, breast cancer, and diabetes. In addition, a group of external collaborators at University of Iowa will participate in studies on microbial pathogens that cause respiratory disease, tularemia, and otitis media, and second group at the Gladstone Institute and Joslin Diabetes Center on metabolic syndrome and diabetes.

Agency
National Institute of Health (NIH)
Institute
Office of The Director, National Institutes of Health (OD)
Type
Biomedical Research Support Shared Instrumentation Grants (S10)
Project #
1S10OD016281-01
Application #
8447837
Study Section
Special Emphasis Panel (ZRG1-BCMB-D (30))
Program Officer
Birken, Steven
Project Start
2013-06-20
Project End
2014-05-31
Budget Start
2013-06-20
Budget End
2014-05-31
Support Year
1
Fiscal Year
2013
Total Cost
$597,064
Indirect Cost
Name
Buck Institute for Age Research
Department
Type
DUNS #
786502351
City
Novato
State
CA
Country
United States
Zip Code
94945
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Meyer, Jesse G; D'Souza, Alexandria K; Sorensen, Dylan J et al. (2016) Quantification of Lysine Acetylation and Succinylation Stoichiometry in Proteins Using Mass Spectrometric Data-Independent Acquisitions (SWATH). J Am Soc Mass Spectrom 27:1758-1771
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Schilling, Birgit; MacLean, Brendan; Held, Jason M et al. (2015) Multiplexed, Scheduled, High-Resolution Parallel Reaction Monitoring on a Full Scan QqTOF Instrument with Integrated Data-Dependent and Targeted Mass Spectrometric Workflows. Anal Chem 87:10222-9
AbouElfetouh, Alaa; Kuhn, Misty L; Hu, Linda I et al. (2015) The E. coli sirtuin CobB shows no preference for enzymatic and nonenzymatic lysine acetylation substrate sites. Microbiologyopen 4:66-83
Schilling, Birgit; Christensen, David; Davis, Robert et al. (2015) Protein acetylation dynamics in response to carbon overflow in Escherichia coli. Mol Microbiol 98:847-63
Nishida, Yuya; Rardin, Matthew J; Carrico, Chris et al. (2015) SIRT5 Regulates both Cytosolic and Mitochondrial Protein Malonylation with Glycolysis as a Major Target. Mol Cell 59:321-32
O'Brien, Robert; DeGiacomo, Francesco; Holcomb, Jennifer et al. (2015) Integration-independent Transgenic Huntington Disease Fragment Mouse Models Reveal Distinct Phenotypes and Life Span in Vivo. J Biol Chem 290:19287-306
French, William R; Zimmerman, Lisa J; Schilling, Birgit et al. (2015) Wavelet-based peak detection and a new charge inference procedure for MS/MS implemented in ProteoWizard's msConvert. J Proteome Res 14:1299-307
Rardin, Matthew J; Schilling, Birgit; Cheng, Lin-Yang et al. (2015) MS1 Peptide Ion Intensity Chromatograms in MS2 (SWATH) Data Independent Acquisitions. Improving Post Acquisition Analysis of Proteomic Experiments. Mol Cell Proteomics 14:2405-19

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