SGER: Solid Support-Assisted Screen of Complex Protein Libraries With Small Molecules

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Intellectual Merit: Few tools are currently available to identify links between proteins and small molecules (1,000 Da, non-biopolymeric), despite the importance of these interactions in many biological processes. To address this problem, a solid support-assisted screen (METSCREEN) of protein libraries with small molecules as a strategy to identify missing steps in biosynthetic pathways, new enzymes that can modify existing metabolites or receptors for small molecules will be developed. METSCREEN consists of linking small molecules to beads and using them to bio-pan phage display or RNA display libraries to identify the proteins that bind to the small molecule. Analysis of the genetic material linked to the proteins allows rapid identification of corresponding genes. As an exciting application of METSCREEN, RNA-protein fusions obtained from RNA display will be used directly to probe DNA chips, permitting rapid qualitative and quantitative identification of possible alternative fates of a pathway intermediate. This will contribute to the rationale design of metabolic engineering strategies. Initially, METSCREEN will be applied towards the identification of Arabidopsis proteins that bind naringenin, a central intermediate in multiple branches of flavonoid biosynthesis. The successful development and implementation of METSCREEN will make a significant contribution to systems biology by filling in difficult-to-establish connections in the genome-proteome-metabolome continuum.Broader Impacts: METSCREEN integrates chemistry and biology in a strategy that, if successful, will permit establishment of direct links between small molecules and the proteins that they bind, filling significant gap in systems biology. The interdisciplinary nature of this project will significantly enrich the training of post-doc, graduate students and undergrads in the PI's lab. This research will continue to enhance the opportunities for the PI to link research and education through the continuous participation of undergraduates in research in the lab, the enrichment of the topic covered in courses taught by the PI (e.g., PB625 Plant Metabolic Engineering) and the delivery of resources to the community for general use (e.g., the Arabidopsis Gene Regulatory Information Server, http://arabidopsis.med.ohio-state.edu).