Generation and Proteome-Wide Screening of Hybrid Combinatorial Libraries

描述 摘要： 随着人类基因组测序和注释的完成，我们现在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不清楚。阐明蛋白质功能的一种方法是找到小分子配体，这些小分子配体专门与感兴趣的蛋白质结合，并扰乱它们的生化和细胞功能。今天化学生物学家面临的一个主要挑战是发现新的小分子探针来寻找新的蛋白质，以便于阐明它们的功能。蛋白质芯片开发的最新进展提供了一个令人兴奋的新机会，首次针对几乎整个人类蛋白质组同时筛选化学文库。然而，由于缺乏通用读数来检测芯片上的配体与蛋白质的结合，利用现有的大多数(如果不是全部)化学文库筛选人类蛋白质芯片是不可行的。我建议借用一个成熟的天然产物的蛋白质结合结构域来作为展示组合肽和非肽文库的支架，以及作为库中每个化合物的嵌入通用标签。这种混合组合文库将适用于使用蛋白质芯片进行蛋白质组范围的筛选，方法是利用来自天然产品的蛋白质结合域与其结合蛋白之间的相互作用，该结合蛋白可以用针对该蛋白质的荧光标记抗体进行检测。此外，已知的蛋白结合域也可以赋予融合配体稳定性和细胞渗透性，增加了来自这种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。如果这项工作取得成功，可能会产生一种新的结构类型的配体，可以用作蛋白质功能的探针。 公共卫生相关性： 这个项目的目的是产生一种新型的能源

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. Public Health Relevance: This project is aimed at generating a new type of h