High-Throughput Assessment of De Novo Protein Design: Generation of Molecular Probes for Guanine Exchange Factors

了解蛋白质相互作用的基本原理有助于设计 具有新功能的蛋白质，这些蛋白质在自然界中仍未被观察到，并为 科学家们将剖析疾病的分子基础。尽管在这方面取得了重大进展 通过在计算机中表达蛋白质来评估计算设计过程 由于与基因合成相关的限制，体外培养一直局限于小样本大小。 寡核苷酸合成领域的最新进展使高通量 用酵母展示筛选设计。我们将利用这项技术来评估新的方法 用于在蛋白质界面包装氨基酸，并确定理想的评分期限目标 设计的模型。在这个过程中，我们将开发出抑制鸟嘌呤交换的蛋白质 对细胞增殖至关重要的因子(GEF)，因此获得了显著的 兴趣作为癌症的治疗靶点。我们还致力于开发新型生物传感器，这些传感器 能够通过将结合位点工程到SnapTag和HaloTag来报告全球环境基金的激活 构造。预计这些结果将对以下领域产生重要的积极影响 蛋白质工程，最终为试剂的发展提供了新的机会 使科学界能够促进我们对生物过程的理解。

Knowledge of the underlying principles of protein-protein interactions facilitates the design of proteins with novel functions that remain unobserved in nature and create new opportunities for scientists to dissect the molecular basis for diseases. Although significant advancements in the field have been made, evaluation of the computational design process by expressing proteins in vitro has been limited to small sample sizes due to constraints associated with gene synthesis. Recent advances in the field of oligonucleotide synthesis have enabled the high-throughput screening of designs with yeast display. We will harness this technology to assess new methods for packing amino acids at protein interfaces and to determine ideal score term targets for designed models. During this process, we will develop proteins that inhibit guanine exchange factors (GEFs), which are critical for cellular proliferation and have thereby garnered significant interest as therapeutic targets for cancer. We also aim to develop novel biosensors that are capable of reporting GEF activation by engineering a binding site into the SnapTag and HaloTag constructs. These results are expected to have an important positive impact for the field of protein engineering, ultimately providing new opportunities for the development of reagents that enable the scientific community to advance our understanding of biological processes.