The Proteomics Shared Resource at Burnham Institute offers a state-of-the-art infrastructure for protein analysis at a global level that is tailored to the specific needs of Cancer Center investigators. The core technologies include methods for upfront sample fractionation and back-end protein identification by mass spectrometry (MS) and database searching. Facility MS instruments include Thermo LTQ and LTQ Orbitrap systems, and Bruker HCTUItra and MALDI Autoflex II TOF/TOF systems. Routine services include protein/peptide molecular weight determination by MALDI-MS, differential 2D-PAGE analysis of low to moderately complex samples, identification of gel separated proteins by tandem MS, and post-translational modification mapping for known peptides and proteins. More recently added services include analysis for complex protein samples by multidimensional liquid chromatography (MD LC-MS/MS), phosphoproteomics using 2D or 3D LC-MS/MS methodology, targeted quantitative proteomics employing isotope-labeling strategies (ICPL/ITRAQ/TM), and differential quantitative analysis of complex samples using label-free strategies. Cancer Center investigators are utilizing these technologies in a variety of ways, but most frequently to identify novel components of protein complexes and protein-protein interactions, to map functionally important post-translational modifications such as phosphorylation and ubiquitylation, and to comprehensively profile quantitative differences in protein abundance in cancer versus normal cells. To facilitate these projects, facility personnel routinely meet and interact with investigators to provide guidance with project design, samples preparation, and data analysis. In addition, the facility entertains a rich portfolio in outreach activities geared toward training investigators and their associates in proteomic technology. The facility's future activities will focus on maintaining high quality services as well as the exploration, testing, and demand-based implementation of novel technologies that will benefit the Center's research programs. Efforts in planning toward these goals include promoting additional technologies for accurate quantitative protein profiling in the low abundance range (SILAC), enhancing data analysis, data management and biological interpretation (pathway analysis), and expanding the facility's personnel and instrumentation capacity to a high resolution, high sensitivity mass spectrometry based proteomics facility. In the past year, 25 Cancer Center members used the Proteomics Shared Resource. The requested CCSG support in the first year of $168,553 is 16.6% of the total projected annual Proteomics Shared Resource budget.
Cancer is thought to arise from the accumulation of a series of somatic mutations. However, the genetic make-up as well as mRNA profiles are incomplete descriptors of the malignant phenotype, since proteins are the ultimate executioners of cellular functions. Proteomics provides complementary analytical solutions for deciphering protein functions that are geared toward a systems-level understanding of the cancer phenotype.
|Gong, Xiao-Min; Ding, Yi; Yu, Jinghua et al. (2015) Structure of the Na,K-ATPase regulatory protein FXYD2b in micelles: implications for membrane-water interfacial arginines. Biochim Biophys Acta 1848:299-306|
|Brun, S N; Markant, S L; Esparza, L A et al. (2015) Survivin as a therapeutic target in Sonic hedgehog-driven medulloblastoma. Oncogene 34:3770-9|
|You, Weon-Kyoo; Yotsumoto, Fusanori; Sakimura, Kenji et al. (2014) NG2 proteoglycan promotes tumor vascularization via integrin-dependent effects on pericyte function. Angiogenesis 17:61-76|
|Vargas, Lina M; Leal, Nancy; Estrada, Lisbell D et al. (2014) EphA4 activation of c-Abl mediates synaptic loss and LTP blockade caused by amyloid-? oligomers. PLoS One 9:e92309|
|Volkmann, Niels; Page, Christopher; Li, Rong et al. (2014) Three-dimensional reconstructions of actin filaments capped by Arp2/3 complex. Eur J Cell Biol 93:179-83|
|Bailey, Ann M; Zhan, Le; Maru, Dipen et al. (2014) FXR silencing in human colon cancer by DNA methylation and KRAS signaling. Am J Physiol Gastrointest Liver Physiol 306:G48-58|
|Valencia, Tania; Kim, Ji Young; Abu-Baker, Shadi et al. (2014) Metabolic reprogramming of stromal fibroblasts through p62-mTORC1 signaling promotes inflammation and tumorigenesis. Cancer Cell 26:121-35|
|Northcott, Paul A; Lee, Catherine; Zichner, Thomas et al. (2014) Enhancer hijacking activates GFI1 family oncogenes in medulloblastoma. Nature 511:428-34|
|Kim, H; Claps, G; Moller, A et al. (2014) Siah2 regulates tight junction integrity and cell polarity through control of ASPP2 stability. Oncogene 33:2004-10|
|Finlay, Darren; Vamos, Mitchell; Gonzalez-Lopez, Marcos et al. (2014) Small-molecule IAP antagonists sensitize cancer cells to TRAIL-induced apoptosis: roles of XIAP and cIAPs. Mol Cancer Ther 13:5-15|
Showing the most recent 10 out of 322 publications