Innovative Technologies for Metabolite Profiling and Natural Products Discovery

自然界的小分子宇宙提供了极其丰富的结构多样性， 理化特性和生物活性。对天然存在的小分子的研究可以 分为两类：1）代谢组学，或研究细胞和组织的小分子含量， 确定（病理）生理上重要的生化途径，和2）天然产物的发现，或 鉴定具有生物活性的化合物。最终，代谢物的研究和 天然产物依赖于能够使它们从复杂的物质中有效分离和富集的方法。 生物样本尽管作出了相当大的努力，但全球战略的丰富和特点， 这些化合物仍然是难以捉摸的。在本申请中，我们建议开发一套创新的 促进任何物理化学类别的小分子富集的技术。K99辅导 阶段：博士后阶段（目标1）的建议将在教授的指导下进行。 Benjamin Cravatt在斯克里普斯研究所。这一阶段的目标是开发一种化学选择性的 标记和富集策略适用于代谢组学分析。该培训将提供以下方面的专门知识： 质谱、结构测定和细胞培养技术。获得的知识将 为目标2和3奠定基础，并过渡到独立的职业生涯。独立ROD阶段： 目标2将侧重于目标1中制定的战略与基于活动的 蛋白质组学和基因组学技术组装有助于细菌生长的生化网络， 坚持不懈一个持久的菌株的医院病原体，铜绿假单胞菌，将进行检查 通过与弗洛伊德罗姆斯伯格教授的合作。AIM3将扩展小分子标记策略 创造一种富集生物活性天然产物的技术。这种方法将有助于检测 天然产品，往往无法使用目前的纯化方法，并使探索 自然界的小分子剧目以前所未有的广度和深度。相关性：该计划具有 通过开发有助于了解 疾病的发病机理，以及治疗剂的发现。我们将把这些方法应用到研究中 铜绿假单胞菌的持久性和致病性以及新型抗菌剂的鉴定。

Nature's small molecule universe offers a remarkably rich assortment of structural diversity, physicochemical characteristics, and bioactivity. The study of naturally occurring small molecules can be divided into two categories: 1) metabolomics, or study of the small molecule content of cells and tissues to identify (patho)physiologically important biochemical pathways, and 2) natural products discovery, or the identification of compounds possessing biological activity. Ultimately, the investigation of metabolites and natural products depends upon methods that enable their effective separation and enrichment from complex biological samples. Despite considerable effort, global strategies for the enrichment and characterization of these compounds have remained elusive. In this application, we propose to develop an innovative set of technologies to facilitate the enrichment of small molecules of any physicochemical class. Mentored K99 phase: The postdoctoral phase (Aim 1) of the proposal will be carried out under the mentorship of Prof. Benjamin Cravatt at The Scripps Research Institute. The goal of this phase is to develop a chemoselective tagging and enrichment strategy applicable to metabolomic profiling. This training will provide expertise in mass spectrometry, structure determination, and cell culture techniques. The acquired knowledge will provide the foundation for Aims 2 and 3 and transition to an independent career. Independent ROD Phase: Aim 2 will focus on application of the strategy developed in Aim 1 in combination with activity-based proteomic and genomic technologies to assemble biochemical networks that contribute to bacterial persistence. A persistent strain of the hospital pathogen, Pseudomonas aeruginosa, will be examined through collaboration with Prof. Floyd Romesberg. Aim 3 will expand on the small-molecule tagging strategy to create a technology for the enrichment of bioactive natural products. This method will facilitate detection of natural products that are often inaccessible using current purification methods and enable exploration of nature's small molecule repertoire with unprecedented scope and depth. Relevance: This program has the potential to improve public health through development of technologies that will assist in understanding disease pathogenesis, as well as discovery of therapeutic agents. We will apply these methods to the study of persistence and pathogenicity in Pseudomonas aeruginosa and the identification of novel antimicrobials.