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

智力优势：目前很少有工具可用于识别蛋白质和小分子（1,000 Da，非生物聚合物）之间的联系，尽管这些相互作用在许多生物过程中很重要。为了解决这一问题，将开发一种固体支持物辅助筛选（METSCREEN）的蛋白质文库与小分子作为识别生物合成途径中缺失步骤的策略，可以修饰现有代谢物或小分子受体的新酶。 METSCREEN包括将小分子连接到珠子上，并将它们用于生物泛噬菌体展示或RNA展示文库，以鉴定与小分子结合的蛋白质。分析与蛋白质相关的遗传物质可以快速识别相应的基因。作为METSCREEN的一个令人兴奋的应用，从RNA展示中获得的RNA-蛋白融合体将直接用于探针DNA芯片，允许快速定性和定量鉴定途径中间体的可能替代命运。 这将有助于代谢工程策略的合理设计。 最初，METSCREEN将被应用于识别结合柚皮素的拟南芥蛋白质，柚皮素是类黄酮生物合成多个分支的中心中间体。METSCREEN的成功开发和实施将填补基因组-蛋白质组-代谢组连续体中难以建立的联系，从而为系统生物学做出重大贡献。更广泛的影响：METSCREEN将化学和生物学整合在一个战略中，如果成功，将允许在小分子和它们结合的蛋白质之间建立直接联系，填补系统生物学中的重大空白。该项目的跨学科性质将大大丰富PI实验室的博士后，研究生和本科生的培训。这项研究将继续提高PI的机会，通过本科生在实验室研究的持续参与，PI教授的课程所涵盖的主题的丰富（例如，PB 625植物代谢工程）和向社区提供资源供一般使用（例如，拟南芥基因调控信息服务器，http://arabidopsis.med.ohio-state.edu）。

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).