Innovative Technologies for Metabolite Profiling and Natural Products Discovery

描述（由申请人提供）：自然界的小分子宇宙提供了非常丰富的结构多样性、物理化学特征和生物活性。对天然存在的小分子的研究可分为两类：（1）代谢组学，或对细胞和组织的小分子含量的研究，以鉴定（病理）生理学上重要的生物化学途径，和（2）天然产物发现，或鉴定具有生物活性的化合物。最终，代谢物和天然产物的研究取决于能够从复杂生物样品中有效分离和富集它们的方法。尽管作出了相当大的努力，但这些化合物的富集和表征的全球战略仍然难以捉摸。在这个应用中，我们建议开发一套创新的技术，以促进任何物理化学类的小分子的富集。 K99阶段：该提案的博士后阶段（目标1）将在斯克里普斯研究所的Benjamin Cravatt教授的指导下进行。这一阶段的目标是开发一种适用于代谢组学分析的化学选择性标记和富集策略。该培训将提供质谱、结构测定和细胞培养技术方面的专业知识。所获得的知识将为目标2和目标3奠定基础，并过渡到独立的职业生涯。 独立R00阶段：目标2将集中于应用目标1中开发的策略，结合基于活性的蛋白质组学和基因组学技术，组装有助于细菌持久性的生化网络。医院病原体铜绿假单胞菌的持续菌株将通过与弗洛伊德罗姆斯伯格教授的合作进行检查。目标3将扩展小分子标记策略，以创建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使用当前纯化方法通常无法获得的天然产物，并能够以前所未有的范围和深度探索自然界的小分子库。 相关性：该计划有可能通过开发有助于了解疾病发病机制的技术以及发现治疗药物来改善公众健康。我们将把这些方法应用于铜绿假单胞菌持久性和致病性的研究以及新型抗菌剂的鉴定。

DESCRIPTION (provided by applicant): 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 physico-chemical 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 chemo-selective 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 R00 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.