This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Two projects are being carried out at the Resource, to provide useful data and to serve as a training platform for Ms. Seo. (1) Characterization Of Vimentin fragments and their post-translational modifications under external stimuli and (2) Characterization Of Fas Associating Factor 1 (FAF 1) and posttranslational modifications in response to heat shock. Methods developed at the BUSM MS Resource for sample preparation and analysis and data interpretation are being applied, with the focus being on MALDI and ESI QoTOF MS and MS/MS and Fourier Transform Ion Cyclotron Mass Spectrometry (FT-ICR-MS). Life phenomena, such as cellular development, proliferation, aging, apoptosis, carcinogenesis, and disease, are regulated by protein differential expression, post-translational modification (PTM) of proteins, protein-protein interactions and so on. PTMs of a protein can determine its activity state, localization, turnover and interactions with other proteins. Despite the importance of PTMs, the study on PTMs has many limitations such as sensitivity, resolution, sequence coverage and the need for high throughput analysis. Especially, we need better resolution for differentiating some kinds of PTMs. Post-translational modifications of Vimentin fragments in HUVECs were globally defined by MALDI-TOF MS and nano-LC-ESI-Q-TOF MS (in Korea). However, these instruments currently do not have the ability to confirm PTMs due to their low accuracy (-50 ppm). Even when immunoblot analysis was employed, difficulties were encountered in confirmation of PTMs, in the cases of some of PTMs such as acetylation, trimethylation, phosphorylation, and sulfonation. At BUSM, the QoTOF MS experiments have been repeated with higher performance instruments, using well-developed methods for the mass spectral analysis and data interpretation. Any still ambiguous details of the characterization of PTMs are being addressed using a home-built 7T FT-ICR-mass spectrometer. Analysis of proteins using FT-ICR-mass spectrometry 1. Characterization of PTMs using MS/MS analysis of post-translational modifications such as phosphorylation, acetylation, oxidation, methylation and sulfonation 2. Methodology development for enrichment of low abundant modified proteins 3. Methodology development for almost 100% peptide sequence coverage of protein for comprehensive understanding. The exact location and diversity of PTMs over the whole peptide sequences will provide some useful information to evaluate the biological effects in the cellular system. This proteomic technique is needed and applied to the biotechnology related industry, pharmaceutical industry and understanding of life phenomena. This research can be used to find out the biomarkers of disease, identify the proteins regulating diseases, diagnose the disease and develop new drugs.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR010888-10
Application #
7369309
Study Section
Special Emphasis Panel (ZRG1-BECM (03))
Project Start
2006-07-01
Project End
2007-06-30
Budget Start
2006-07-01
Budget End
2007-06-30
Support Year
10
Fiscal Year
2006
Total Cost
$9,467
Indirect Cost
Name
Boston University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code
02118
Lu, Yanyan; Jiang, Yan; Prokaeva, Tatiana et al. (2017) Oxidative Post-Translational Modifications of an Amyloidogenic Immunoglobulin Light Chain Protein. Int J Mass Spectrom 416:71-79
Sethi, Manveen K; Zaia, Joseph (2017) Extracellular matrix proteomics in schizophrenia and Alzheimer's disease. Anal Bioanal Chem 409:379-394
Hu, Han; Khatri, Kshitij; Zaia, Joseph (2017) Algorithms and design strategies towards automated glycoproteomics analysis. Mass Spectrom Rev 36:475-498
Ji, Yuhuan; Bachschmid, Markus M; Costello, Catherine E et al. (2016) S- to N-Palmitoyl Transfer During Proteomic Sample Preparation. J Am Soc Mass Spectrom 27:677-85
Hu, Han; Khatri, Kshitij; Klein, Joshua et al. (2016) A review of methods for interpretation of glycopeptide tandem mass spectral data. Glycoconj J 33:285-96
Pu, Yi; Ridgeway, Mark E; Glaskin, Rebecca S et al. (2016) Separation and Identification of Isomeric Glycans by Selected Accumulation-Trapped Ion Mobility Spectrometry-Electron Activated Dissociation Tandem Mass Spectrometry. Anal Chem 88:3440-3
Wang, Yun Hwa Walter; Meyer, Rosana D; Bondzie, Philip A et al. (2016) IGPR-1 Is Required for Endothelial Cell-Cell Adhesion and Barrier Function. J Mol Biol 428:5019-5033
Srinivasan, Srimathi; Chitalia, Vipul; Meyer, Rosana D et al. (2015) Hypoxia-induced expression of phosducin-like 3 regulates expression of VEGFR-2 and promotes angiogenesis. Angiogenesis 18:449-62
Yu, Xiang; Sargaeva, Nadezda P; Thompson, Christopher J et al. (2015) In-Source Decay Characterization of Isoaspartate and ?-Peptides. Int J Mass Spectrom 390:101-109
Steinhorn, Benjamin S; Loscalzo, Joseph; Michel, Thomas (2015) Nitroglycerin and Nitric Oxide--A Rondo of Themes in Cardiovascular Therapeutics. N Engl J Med 373:277-80

Showing the most recent 10 out of 253 publications