CAREER: Teflon Proteins for Protein-Protein Interaction Ligand Discovery

美国国家科学基金会化学部的生命过程化学计划资助明尼苏达大学的威廉·波梅兰茨博士开发氟检测的核磁共振方法，以研究蛋白质之间的相互作用。波美兰茨实验室的研究和教育目标解决了蛋白质之间生物分子通信的长期问题，借鉴了化学和生物学学科的研究结果。被称为转录因子的蛋白质是细胞功能的主要调节因子。它们促进了复杂的信息传输，这些信息指定了DNA代码是如何用于制造蛋白质的。这项研究试图了解蛋白质与其潜在生物学之间相互作用的细节。氟元素将被用作蛋白质界面上难以检测到的相互作用的敏感报告。考虑到氟的独特光谱性质和天然生物分子中不存在氟，这种方法是有利的。鉴于转录因子的重要性，这种以氟为基础的研究蛋白质-配体相互作用的方法可以显著增加新的蛋白质靶标的研究，从而为提高对各种蛋白质功能的理解开辟新的途径。该项目将包括对没有化学/生物界面工作经验的学生进行培训，并将使Osher终身学习社区对科学素养和化学生物学在科学和技术中的作用感兴趣。这项研究的重点是开发氟核磁共振(核磁共振)方法来检测、量化和定义转录因子-蛋白质界面上包括表观遗传调节蛋白在内的新的相互作用模式。由于芳香族氨基酸在蛋白质结合部位的富集性，为了报道蛋白质结合作用，将评估多种用于序列选择性掺入含氟芳香族氨基酸的标记策略。氟核对化学环境变化的高响应性导致了简化的一维生物分子核磁共振谱，可以快速获得并易于解释。这种方法将与已知的配体进行评估，以表征它们的相互作用模式，并使用小分子文库以发现的形式进行评估。这些研究将扩展到通过细胞内氟-19核磁共振获取生理条件下的机制信息。细胞摄取氟标记的多肽的起源将使用稀土移位试剂来区分胞外多肽和内化多肽。这种氟-19核磁共振方法将成为一种基于结构的工具，为寻找挑战蛋白质-蛋白质相互作用的新化学探针奠定基础。鉴于转录因子的重要性，所提出的研究蛋白质-配体相互作用的~(19)F核磁共振方法可以显著增加新的蛋白质靶标的库，为提高对其生物学的理解开辟了途径。

The Chemistry of Life Processes Program in the Chemistry Division of NSF is funding Dr. William Pomerantz from the University of Minnesota to develop nuclear magnetic resonance methods of fluorine detection to study interactions between proteins. The research and educational goals in the Pomerantz lab address long-standing questions about biomolecular communications between proteins, drawing from findings in the disciplines of both chemistry and biology. Proteins termed transcription factors are the master regulators of cellular function. They facilitate the complicated transfer of information that specifies how the DNA code is used to make proteins. This research seeks to understand the details of the interactions between proteins and their underlying biology. The element fluorine will be used as a sensitive reporter for difficult-to-detect interactions at protein interfaces. This method is advantageous in view of fluorine's unique spectroscopic properties and its absence in natural biological molecules. Given the importance of transcription factors, this fluorine-based approach for studying protein-ligand interactions could significantly increase the repertoire of new protein targets for study and thereby open up avenues for improving the understanding of a wide variety of protein function. This project will include training for students with no prior experience of working at the chemistry/biology interface and will engage the Osher Life-Long Learning community's interests in science literacy and the role of chemical biology in science and technology. This research project is focused on the development of fluorine nuclear magnetic resonance spectroscopy (NMR) methods for detecting, quantifying, and defining novel modes of interactions at transcription factor-protein interfaces that include epigenetic regulatory proteins. Due to the enrichment of aromatic amino acids at protein-binding sites, multiple labeling strategies for sequence selective incorporation of fluorinated aromatic amino acids will be evaluated for reporting on protein binding interactions. The hyper-responsiveness of the fluorine nucleus to changes in chemical environment leads to simplified 1D-biomolecular NMR spectra that can be rapidly acquired and readily interpreted. This method will be evaluated against known ligands to characterize their mode of interaction and also in discovery format using small molecule libraries. These studies will be extended to acquiring mechanistic information under physiological conditions by in-cell fluorine-19 NMR. The origins of cellular uptake of fluorine-labeled peptide macrocylces will be investigated using lanthanide shift reagents for differentiating extracellular versus internalized peptides. This fluorine-19 NMR method will set the stage for use as a structure-based tool to find new chemical probes for challenging protein-protein interactions. Given the importance of transcription factors, the proposed fluorine-19 NMR approach to study protein-ligand interactions could significantly increase the repertoire of new protein targets opening up avenues for improving the understanding of their biology.