Core B will provide proteomics and protein analysis services for PPG projects. Throughout the last funding cycle, the protein analysis component of Core B has significantly expanded to meet the needs of the individual projects in the PPG. In addition to mass spectrometry (MS)-based footprinting technology for the mapping of protein-protein and protein-nucleic acid interactions proposed in the previous application, we have added the following new services to the Core: 1) the AKTA-fast protein liquid chromatography (FPLC), which has played a critical role in obtaining purified recombinant proteins for biochemical/biophysical characterization for former Project 3 and antibody production for former Project 2;and 2) the EnSpire multimode plate reader to provide a new service for highly sensitive, rapid and robust monitoring of protein-protein and protein-nucleic acid interactions. MS-based proteomic approaches enabled us to identify new protein-protein interactions and posttranslational modifications, and have thus been instrumental for advancing the research of former Projects 1 and 2, respectively. The above technologies will continue to play an important role in accomplishing the research objectives of the PPG projects proposed in this competitive renewal. In particular, Specific Aims of the Proteomics and Protein Analysis Core are:
Aim 1 : to perform MS-based protein footprinting experiments to identify contact amino acids in the context of large biologically relevant nucleoprotein and protein-protein complexes;
Aim 2 : to develop novel applications of the EnSpire multimode plate reader for biochemical characterization of protein-nucleic acid and protein-protein interactions using homogeneous time-resolved fluorescence energy transfer (HTRF), AlphaLISA, and AlphaScreen (ALPHA) based technologies;
Aim 3 : to provide a column chromatography service using AKTA-FPLC to assist individual PPG projects to isolate low abundant native protein-protein complexes from mammalian cells and purify mg quantities of poorly soluble recombinant proteins from E. coli for their subsequent biochemical and biophysical characterization;
Aim 4 : to conduct MS-based proteomics experiments to identify interacting partners and protein posttranslational modifications. Taken together, Core B will provide the wide range of protein analysis services. Importantly, these services will be utilized by all four PPG projects.

Public Health Relevance

Overall goals of the PPG include delineating mechanisms of T-cell leukemia induced by the human T cell leukemia virus type 1 (HTLV-1). These studies require state-of-the-art proteomics and protein analysis technologies, which will be provided by Core B.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
2P01CA100730-11A1
Application #
8742037
Study Section
Special Emphasis Panel (ZCA1-RPRB-J (M1))
Project Start
2014-09-23
Project End
2019-08-31
Budget Start
2014-09-23
Budget End
2015-08-31
Support Year
11
Fiscal Year
2014
Total Cost
$176,110
Indirect Cost
$46,374
Name
Ohio State University
Department
Type
DUNS #
832127323
City
Columbus
State
OH
Country
United States
Zip Code
43210
Webb, Lindsay M; Amici, Stephanie A; Jablonski, Kyle A et al. (2017) PRMT5-Selective Inhibitors Suppress Inflammatory T Cell Responses and Experimental Autoimmune Encephalomyelitis. J Immunol 198:1439-1451
Esser, Alison K; Rauch, Daniel A; Xiang, Jingyu et al. (2017) HTLV-1 viral oncogene HBZ induces osteolytic bone disease in transgenic mice. Oncotarget 8:69250-69263
Al-Saleem, Jacob; Kvaratskhelia, Mamuka; Green, Patrick L (2017) Methods for Identifying and Examining HTLV-1 HBZ Post-translational Modifications. Methods Mol Biol 1582:111-126
Fontana, Francesca; Ge, Xia; Su, Xinming et al. (2017) Evaluating Acetate Metabolism for Imaging and Targeting in Multiple Myeloma. Clin Cancer Res 23:416-429
Ratner, L; Rauch, D; Abel, H et al. (2016) Dose-adjusted EPOCH chemotherapy with bortezomib and raltegravir for human T-cell leukemia virus-associated adult T-cell leukemia lymphoma. Blood Cancer J 6:e408
Panfil, Amanda R; Martinez, Michael P; Ratner, Lee et al. (2016) Human T-cell leukemia virus-associated malignancy. Curr Opin Virol 20:40-46
Kawatsuki, A; Yasunaga, J-I; Mitobe, Y et al. (2016) HTLV-1 bZIP factor protein targets the Rb/E2F-1 pathway to promote proliferation and apoptosis of primary CD4(+) T cells. Oncogene 35:4509-17
Niewiesk, Stefan (2016) Animals Models of Human T Cell Leukemia Virus Type I Leukemogenesis. ILAR J 57:3-11
Su, Xinming; Esser, Alison K; Amend, Sarah R et al. (2016) Antagonizing Integrin ?3 Increases Immunosuppression in Cancer. Cancer Res 76:3484-95
Singh, Deepali; Boeras, Ioana; Singh, Gatikrushna et al. (2016) Isolation of Cognate Cellular and Viral Ribonucleoprotein Complexes of HIV-1 RNA Applicable to Proteomic Discovery and Molecular Investigations. Methods Mol Biol 1354:133-46

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