This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Metabolomics is the study of the repertoire of non-proteinaceous, endogenously-synthesized small molecules present in any organism. Small molecules present in the metabolome are ultimately the product of cellular metabolism. These molecules are varied from peptide, glucose, cholesterol, ATP and lipids etc. The change in levels of these metabolites in consequence of certain metabolic process or disease state and can be used as biomarkers. The power of metabolomic investigations versus transcriptomic and proteomic ones is that the metabolite dictionary is believed to be an order of magnitude smaller than the repertoire of expressed genes in an organism. In addition, metabolite profiles provide direct versus inferred readouts on the functional consequences of genetic and environmental changes. Furthermore, small metabolites structure is independent of the species under investigation and can be easily relate to different species. Recent developments at the University of Nebraska have led to the advent of genomics, bioinformatics and proteomics supported by dedication of faculty lines and creation of related core facilities at both UNL and UNMC. Therefore, we have developed a new capacity in the complementary area of metabolomics, which is at the cutting edge of functional biology research and is important for the programs of a number of investigators in the Redox Biology Center as well as for the other related research labs at UNL. Mass spectrometry based high-throughput proteomics and metabolomics, a part of system biology, is foundational for a full understanding of a biological system including a metabolic process or disease state, and has received considerable attention since the sequencing of the human genome. Oxidative/nitrosative damage by reactive radical species appears to be central to the pathogenesis of many human diseases and is central to a broad range of biotic and abiotic stress and physiological responses in microorganisms, plants and other animals. Specific modified proteins or metabolites are generated following stressing insults and accumulate in different degenerated tissues and fluids, determining altered organ functionalities. System biology based technologies have been propitious to the development of molecular medicine, especially in the discovery of diagnostic biomarkers of oxidative and nitrosative stress enabling early detection of diseases, such as lung cancer, from these biological fluids. The Laboratory for Proteomics and Metabolomics analysis at the Redox Biology Center (RBC) supports scientists within the RBC and the University of Nebraska. The Core provides all the tools of modern functional proteomics and metabolomics. Equipped with cutting edge mass spectrometry based state-of-the-art technologies for proteomics and metabolomics;protein and metabolite profiling, protein identification, protein and peptide fractionation and quantitation, differential expression analysis, shot gun proteomics, personalized experimental design consultation and comprehensive individualized bioinformatics support. The past year was very productive for the core. We were able to implement the only discovery proteomic differential expression analysis platform (mudPIT) in the state of Nebraska and were able to provide service to 27 users in 2010 and 35 in 2011. We implemented a state of the art proteomic bioinformatic platform for extensive data analysis. In addition, a new SRM/MRM targeted proteome analytical platform was developed and used for service at UNL and UNMC researchers. At present this is the only platform we have to achieve the targeted proteome analysis and validation. Late last year we procured a UPLC system and set up a routine metabolite profiling platform. Since there is no centralized metabolite library available to search and identify we developed new Bioinformatic software called METLIBUNL. Using this software one can add many new metabolite to the library and search the data. This unit is available for all users. We also procured other innovative software such as the LIPDVIEW for lipid profiling. The utilization of the unique technology and techniques available at the Laboratory for Proteomics and Metabolomics provided data that supported many investigators with the submission of several external grants including RO1 at NIH. This unit also attracted two industries including BioRad for a product development and testing.
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