Generation and Proteome-Wide Screening of Hybrid Combinatorial Libraries

混合组合文库的生成和全蛋白质组筛选

• 批准号: 8705479
• 负责人: Jun O. Liu
• 金额: $ 81.18万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8705479
• 关键词: Antibodies; Binding; Binding Proteins; Biochemical; Biological Factors; Cataloging; Catalogs; Cell physiology; Cells; Chemicals; Development; Generations; Genome; Human; Human Genome; Hybrids; Label; Lead; Libraries; Ligands; Peptide Library; Permeability; Pharmaceutical Preparations; Probability; Protein Binding Domain; Protein Microchips; Proteins; Proteome; Time; Work; abstracting; combinatorial; interest; protein function; public health relevance; scaffold; screening; small molecule; small molecule libraries

DESCRIPTION Abstract: With the completion of the sequencing and annotation of the human genome, we now have a complete catalog of all human proteins encoded in the genome. However, the functions of a majority of the proteins remain unknown. One way to elucidate the functions of proteins is to find small molecule ligands that specifically bind to the proteins of interest and perturb their biochemical and cellular functions. A major challenge for chemical biologists today is to discover new small molecule probes for new proteins to facilitate the elucidation of their functions. The recent advance in the development of protein chips has offered an exciting new opportunity to simultaneously screen chemical libraries against nearly the entire human proteome for the first time. However, screening of the human protein chips is not yet feasible with most, if not all, existing chemical libraries due to the lack of a universal readout for detecting the binding of a ligand to a protein on chips. I propose to borrow a well-established protein-binding domain of natural products to serve both as a scaffold to present combinatorial peptide and non-peptide libraries and as an embedded universal tag for each compound in the library. Such hybrid combinatorial libraries will be amenable to proteome-wide screening using protein chips by exploiting the interaction between the protein binding domain derived from natural products and its binding protein, which can be detected with a fluorescently labeled antibody against the protein. In addition, known protein-binding domain can also confer stability and cell permeability to the fused ligands, increasing the probability that hits from such hybrid combinatorial libraries will be readily applicable to study the cellular functions of the relevant target proteins. This work, if successful, may lead to a new structural class of ligands that can be used as probes of protein function.

描述 抽象的： 随着人类基因组测序和注释的完成，我们现在拥有了基因组中编码的所有人类蛋白质的完整目录。然而，大多数蛋白质的功能仍然未知。阐明蛋白质功能的一种方法是找到与感兴趣的蛋白质特异性结合并扰乱其生化和细胞功能的小分子配体。当今化学生物学家面临的主要挑战是发现新蛋白质的新小分子探针，以促进阐明其功能。蛋白质芯片开发的最新进展为首次针对几乎整个人类蛋白质组同时筛选化学文库提供了令人兴奋的新机会。然而，由于缺乏用于检测配体与芯片上蛋白质结合的通用读数，对于大多数（如果不是全部）现有化学库来说，人类蛋白质芯片的筛选尚不可行。我建议借用天然产物的成熟蛋白质结合域，既作为呈现组合肽和非肽库的支架，又作为库中每种化合物的嵌入式通用标签。这种混合组合文库将适合使用蛋白质芯片进行蛋白质组范围的筛选，通过利用源自天然产物的蛋白质结合结构域与其结合蛋白之间的相互作用，可以用针对该蛋白质的荧光标记抗体来检测。此外，已知的蛋白质结合结构域还可以赋予融合配体稳定性和细胞通透性，从而增加来自此类混合组合文库的命中将很容易应用于研究相关靶蛋白的细胞功能的可能性。这项工作如果成功，可能会产生一类新的配体结构，可用作蛋白质功能的探针。