The Interactions Core: Mega-sequencing, Proteomics, Informatics and Nitrosylation will provide proteomic analysis, mRNA mega-sequencing, bioinformatic analyses, and biostatistics to support the research projects. For proteomic analysis of antioxidant, anti-inflammatory, NADPH oxidase, hedgehog, and estrogen signaling pathway targets, affecting protein expression levels and redox-based protein post-translational modifications (PTM), particularly S-nitrosylation on specific protein cysteine thiols, we will collaborate with MU's Charles W. Gehrke Proteomics Center to develop an integrative proteomics strategy including both gel- and LC/MSbased quantitative proteomics. We will develop protein cysteine post-translational modification PTM site mapping using CID/ETD technology and provide our expertise to identify potential redox targets and aid in mechanistic studies. High-throughput mRNA mega-sequencing will also be completed by personnel in this core. We will work with the MU DNA Facility and its ultra-high throughput sequencing platform. Sequence data output will be routed directly to the bioinformatician for initial processing, library construction, and screening. In later years of the funding period, we will develop protocols to look at protein-level changes in plasma. Both genomic and proteomic data will be integrated into a pathway interaction model that will provide hypotheses for experimental validation. Overall the Core will provide support to the research projects and work to develop a deeper understanding of how botanicals can influence signaling pathways.
In order to understand signaling pathways and how they are influenced by botanicals, it is necessary to examine gene- and protein-level information. Based on this information, a pathway model will be constructed and validated so that Center scientists can have a fundamental understanding of how botanicals affect the proposed disease models. By helping to provide this information, the Interactions Core will enhance the value of all research projects.
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