? This proposal requests funds for a Thermoelectron (Sunnyvale, CA) model LTQ FT22000 hybrid linear ion trap/Fourier transform mass spectrometer, equipped with an electrospray ionization (ESI) source, infrared multiphoton dissociation (IRMPD) and electron capture dissociation (ECD). The instrument would directly benefit a group of major users all of whom have NIH research support and all of whom currently use mass spectrometry in their research. Many of the investigators are participants in one or more NIH-supported roadmap and program project grants, which incorporate mass spectrometric analysis as an integral part of the research effort and provide a means for expanding the number of investigators benefiting from these resources. This includes the Johns Hopkins Center for Proteomics of Ischemia and Hypoxia (N01 HV28180, supported by NHLBI) and the Johns Hopkins Technology Center for Networks and Pathways of Lysine Modifications (TCNP U54 RR020839, supported by NIH). All of the features and capabilities of this instrument, which distinguish it from those available on current instruments at Johns Hopkins, will be utilized. In work by several investigators on lysine modifications and the structures of histones, high mass resolution (50,000 to 100,000) will distinguish trimethyl and acetylation sites (which may exist at the same site on different molecules). ECD and IRMPD will be used to fragment the very large branched peptide structures resulting from SUMOylation, as well as very large, partially digested peptide fragments needed to map polyacetylated lysines in histones and histone transferases. ECD will also be critical for the structural analysis of carbohydrates, O-linked glycopeptides and negatively charged post- translational modifications, primarily phosphorylation. Many of the studies here are proteomics studies, utilizing protein databases for identification of proteins in complex mixtures (as well as their PTMs) from their digested peptides. High mass accuracy (1-5 ppm) is key to providing correct identifications. The instrument will be housed in and managed by the Middle Atlantic Mass Spectrometry Laboratory and operated by four individuals representing the Major Users and collaborators. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biomedical Research Support Shared Instrumentation Grants (S10)
Project #
1S10RR023025-01
Application #
7125654
Study Section
Special Emphasis Panel (ZRG1-BCMB-D (30))
Program Officer
Tingle, Marjorie
Project Start
2007-05-01
Project End
2008-04-30
Budget Start
2007-05-01
Budget End
2008-04-30
Support Year
1
Fiscal Year
2007
Total Cost
$928,365
Indirect Cost
Name
Johns Hopkins University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Jhaveri, Darshil T; Kim, Min-Sik; Thompson, Elizabeth D et al. (2016) Using Quantitative Seroproteomics to Identify Antibody Biomarkers in Pancreatic Cancer. Cancer Immunol Res 4:225-33
Zhong, Jun; Martinez, Marissa; Sengupta, Srona et al. (2015) Quantitative phosphoproteomics reveals crosstalk between phosphorylation and O-GlcNAc in the DNA damage response pathway. Proteomics 15:591-607
Fiedler, Katherine L; Bheda, Poonam; Dai, Junbiao et al. (2013) A quantitative analysis of histone methylation and acetylation isoforms from their deuteroacetylated derivatives: application to a series of knockout mutants. J Mass Spectrom 48:608-15
Fiedler, Katherine L; Cotter, Robert J (2013) Using glycinylation, a chemical derivatization technique, for the quantitation of ubiquitinated proteins. Anal Chem 85:5827-34
Hansen, Anne-Marie; Chaerkady, Raghothama; Sharma, Jyoti et al. (2013) The Escherichia coli phosphotyrosine proteome relates to core pathways and virulence. PLoS Pathog 9:e1003403
Kim, Min-Sik; Pandey, Akhilesh (2012) Electron transfer dissociation mass spectrometry in proteomics. Proteomics 12:530-42
Osula, Omoruyi; Swatkoski, Stephen; Cotter, Robert J (2012) Identification of protein SUMOylation sites by mass spectrometry using combined microwave-assisted aspartic acid cleavage and tryptic digestion. J Mass Spectrom 47:644-54
van Bodegom, Diederik; Zhong, Jun; Kopp, Nadja et al. (2012) Differences in signaling through the B-cell leukemia oncoprotein CRLF2 in response to TSLP and through mutant JAK2. Blood 120:2853-63
Hersman, Elisabeth; Nelson, Dwella M; Griffith, Wendell P et al. (2012) Analysis of Histone Modifications from Tryptic Peptides of Deuteroacetylated Isoforms. Int J Mass Spectrom 312:5-16
Zhong, Jun; Kim, Min-Sik; Chaerkady, Raghothama et al. (2012) TSLP signaling network revealed by SILAC-based phosphoproteomics. Mol Cell Proteomics 11:M112.017764

Showing the most recent 10 out of 18 publications