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项目资助明尼苏达大学的William Pomerantz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。Pomerantz实验室的研究和教育目标是解决长期存在的关于蛋白质之间生物分子通信的问题，从化学和生物学的学科发现中汲取灵感。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递，这些信息指明了DNA密码是如何用来制造蛋白质的。这项研究旨在了解蛋白质之间相互作用的细节及其潜在的生物学。氟元素将被用作难以检测的蛋白质界面相互作用的敏感报告。由于氟具有独特的光谱特性，且在天然生物分子中不存在，因此该方法具有优势。鉴于转录因子的重要性，这种基于氟的研究蛋白质-配体相互作用的方法可以显著增加新的蛋白质靶点的研究库，从而为提高对各种蛋白质功能的理解开辟了途径。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，并将吸引Osher终身学习社区对科学素养和化学生物学在科学技术中的作用的兴趣。该研究项目的重点是氟核磁共振波谱（NMR）方法的发展，用于检测、量化和定义转录因子-蛋白质界面（包括表观遗传调节蛋白）相互作用的新模式。由于芳香族氨基酸在蛋白质结合位点的富集，将对序列选择性结合含氟芳香族氨基酸的多种标记策略进行评估，以报道蛋白质结合相互作用。氟核对化学环境变化的高响应性导致了简化的1d生物分子核磁共振光谱，可以快速获得并易于解释。该方法将对已知配体进行评估，以表征其相互作用模式，并使用小分子文库进行发现格式。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。细胞摄取氟标记肽大环的起源将使用镧系移位试剂进行研究，以区分细胞外肽和内化肽。这种氟-19核磁共振方法将作为一种基于结构的工具，为寻找新的化学探针来挑战蛋白质-蛋白质相互作用奠定基础。鉴于转录因子的重要性，提出的氟-19核磁共振方法来研究蛋白质-配体相互作用可以显著增加新的蛋白质靶点的曲目，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.