A Genetically Programmable Module for Screening of Protease Inhibitors based on Quantum Dot-Donor Fluorescence Resonance Energy Transfer

基于量子点供体荧光共振能量转移的可遗传编程的蛋白酶抑制剂筛选模块加州大学河滨分校的Wilfred Chen-0755775智力优点这一合作项目的总体目标是开发一种易于适应的遗传可编程模块，用于筛选多种蛋白酶的抑制剂。该方法是产生一个量子点(QD)修饰的，可用作荧光共振能量转移(FRET)底物的蛋白酶特异性蛋白质模块，用于探测蛋白酶活性。这将导致新一代可遗传可编程蛋白质模块作为基于QD的FRET底物，用于发现蛋白酶抑制剂。高通量筛选分析(HTS)将使用这些新的基于QD的FRET底物，这些底物可以通过穿透细胞的TAT多肽在细胞内传递。这一结合将使首席研究人员能够以HTS的方式提供对蛋白酶活性和相应抑制剂有效性的快速而灵敏的评估。广泛的影响：可调生物分子的开发与基于FRET的HTS筛选的集成代表着一项独特的努力，通过药物发现的实施扩展了蛋白质工程的基础发展。参与这项研究的研究生和博士后研究人员将获得连接生物化学、现代遗传学和高温超导的重要接口和协同作用的综合视角。

A Genetically Programmable Module for Screening of Protease Inhibitors Based on Quantum Dot-Donor Fluorescence Resonance Energy TransferWilfred Chen University of California-RiversideCBET-0755775Intellectual Merit The overall objective of this collaborative project is to develop a genetically programmable module that is easily adaptable for screening inhibitors for a wide range of proteases. The approach is to generate a quantum dot (QD)-modified, protease-specific protein module that can be used as a Fluorescence Resonance Energy Transfer (FRET) substrate for probing protease activity. This will result in a new generation of genetically programmable protein modules as QD-based FRET substrates for protease inhibitor discovery. High-throughput screening analysis (HTS) of protease activity will be achieved using these new QD-based FRET substrates that can be delivered intracellularly by a cell-penetrating TAT peptide. The combination will enable the Principal Investigators to provide a rapid and sensitive assessment of protease activity and the corresponding inhibitor efficacies in a HTS manner.Broader Impact: The integration of the development of tunable biomolecules with FRET-based HTS screening represents a unique effort that expands the fundamental development of protein engineering with the implementation of drug discovery. Graduate students and postdoctoral researchers participating in this research will gain an integrated perspective of the important interfaces and synergies connecting biochemistry, modern genetics, and HTS.