Chemical Tools for Precision Metabolic Labeling and Detection of Terpenes and Prenylated Molecules

用于萜烯和异戊二烯化分子的精密代谢标记和检测的化学工具

• 批准号: 2204170
• 负责人: Joshua Baccile
• 金额: $ 43.68万
• 依托单位: University of Tennessee Knoxville
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2204170&HistoricalAwards=false
• 关键词: Chemical; Tools; Precision; Metabolic; Labeling

With the support of the Chemistry of Life Processes (CLP) Program in the Division of Chemistry, Joshua Baccile of the University of Tennessee is studying two biologically important classes of chemical entities, terpenes and prenylated molecules. They are among the largest classes of biomolecules and are both critical to basic life processes and clinically important as drugs. Terpenes and prenylated molecules both derive in part or in full, from the two structurally related five-carbon building blocks, isopentenyl pyrophosphate and dimethylallyl pyrophosphate (IPP and DMAPP, respectively). IPP and DMAPP are therefore among the most significant metabolic precursors in biology. Despite this prominence, chemical tools and strategies to study IPP and DMAPP in cells are lacking. Dr. Baccile aims to bridge this scientific gap through the development of a suite of IPP- and DMAPP-based chemical probes and methods for the metabolic labeling of terpenes and prenylated molecules and their detection in a cell-specific manner. The proposed tools and experiments will also enable a systematic evaluation of the roles of IPP and DMAPP themselves, which is not currently possible. In addition to the chemical innovation, Dr. Baccile will integrate objectives for innovation in student research mentorship, teaching, and learning through the introduction of new pedagogical training methods in the laboratory. Baccile will endeavor to broaden participation in chemistry through a dedicated summer program for underrepresented students in STEM (science, technology, engineering and mathematics) from community colleges.This research project sets out to investigate a facile and modular synthesis of IPP and DMAPP probes, which leverages a “prodrug”-like mechanism for precision metabolic labeling. While related strategies have been employed for metabolic labeling of glycans and proteins, this approach is innovative in natural product and post-translational modification (PTM) research. The proposed cell-type specific isotopic labeling and enrichment methods will enable detailed studies of the isoprenoid pathway in heterogenous cell populations, which could be transformative in our understanding of host-microbe interactions and tumor development. Currently, five-carbon prenyl PTMs are presumed to be absent in life in contrast to the well-established higher order protein prenylations known, specifically protein farnesylation (15 carbons) and geranylgeranylation (20 carbons). However, chemical probes engineered to definitively rule out such five-carbon modifications have not previously been tested. Successful application of the proposed probes will enable the potential identification of five-carbon prenyl modifications, which would be a paradigm shift in the field of protein prenylation. Significant regulatory mechanisms and signaling cascades could be mediated through this yet unknown PTM. We note that the proposed synthetic approach is modular and readily adapted to other pyrophosphate metabolites. This project is an important step toward the long-term goal of creating resources to study the biosynthesis, regulation, and function of biomolecules, with a focus on using isotopic labeling and enrichment chemistry to understand interkingdom signaling.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学系生命过程化学(CLP)项目的支持下，田纳西大学的Joshua Baccile正在研究两类具有生物重要性的化学实体--萜烯和戊烯分子。它们是最大的生物分子之一，既是基本生命过程的关键，也是临床上重要的药物。萜类和戊烯基化分子都部分或全部来自两个结构相关的五碳构筑块，即异戊烯基焦磷酸酯和二甲基烯丙基焦磷酸酯(分别为IPP和DMAPP)。因此，IPP和DMAPP是生物学中最重要的代谢前体。尽管这一点很突出，但缺乏研究细胞中IPP和DMAPP的化学工具和策略。Baccile博士的目标是通过开发一套基于IPP和DMAPP的化学探针和方法来弥合这一科学鸿沟，以代谢标记萜类和异戊烯分子，并以细胞特异性的方式检测它们。拟议的工具和实验还将使人们能够系统地评价IPP和DMAPP本身的作用，这在目前是不可能的。除了化学创新，Baccile博士还将通过在实验室引入新的教学培训方法来整合学生研究指导、教学和学习方面的创新目标。Baccile将努力通过一个专门的暑期项目，为来自社区大学的STEM(科学、技术、工程和数学)中未被充分代表的学生提供一个专门的暑期项目，以扩大对化学的参与。这个研究项目旨在研究一种简便和模块化的IPP和DMAPP探针的合成，它利用一种类似“前药物”的机制进行精确的代谢标记。虽然相关的策略已被用于糖和蛋白质的代谢标记，但这种方法在天然产物和翻译后修饰(PTM)研究中是创新的。所提出的细胞型特异性同位素标记和浓缩方法将使我们能够详细研究异种细胞群体中的异戊二烯途径，这可能对我们理解宿主-微生物相互作用和肿瘤发展具有革命性意义。目前，五碳戊烯基PTM被认为在生命中是不存在的，这与已知的已知的高阶蛋白质预烯基化形成鲜明对比，特别是蛋白质的法尼化(15个碳)和香叶基香叶化(20个碳)。然而，设计用来明确排除这种五碳修饰的化学探针以前从未进行过测试。所提出的探针的成功应用将使识别五碳戊烯基修饰成为可能，这将是蛋白质戊烯基化领域的一种范式转变。重要的调控机制和信号级联反应可以通过这种未知的PTM来调节。我们注意到，建议的合成方法是模块化的，很容易适应其他焦磷酸盐代谢物。这个项目是朝着创造资源来研究生物分子的生物合成、调节和功能的长期目标迈出的重要一步，重点是使用同位素标记和浓缩化学来理解跨界信号。该奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Membrane Permeant Analogs for Independent Cellular Introduction of the Terpene Precursors Isopentenyl‐ and Dimethylallyl‐Pyrophosphate

用于独立细胞引入萜烯前体异戊烯基和二甲基烯丙基焦磷酸盐的膜渗透类似物

• DOI: 10.1002/cbic.202200512
• 发表时间: 2023
• 期刊: ChemBioChem
• 影响因子: 3.2
• 作者: Rossi, Francis M.;McBee, Dillon P.;Trybala, Thomas N.;Hulsey, Zackary N.;Gonzalez Curbelo, Camila;Mazur, William;Baccile, Joshua A.
• 通讯作者: Baccile, Joshua A.