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蛋白酶抑制剂筛选基因可编程模块wilfred Chen加利福尼亚大学河滨分校这个合作项目的总体目标是开发一个基因可编程模块，它可以很容易地适用于筛选多种蛋白酶抑制剂。该方法是生成量子点（QD）修饰的蛋白酶特异性蛋白质模块，该模块可用作荧光共振能量转移（FRET）底物，用于探测蛋白酶活性。这将导致新一代基因可编程蛋白模块作为基于量子点的FRET底物，用于蛋白酶抑制剂的发现。蛋白酶活性的高通量筛选分析（HTS）将使用这些新的基于量子点的FRET底物，这些底物可以通过穿透细胞的TAT肽在细胞内传递。该组合将使主要研究人员能够以HTS方式快速、灵敏地评估蛋白酶活性和相应抑制剂的有效性。更广泛的影响：可调生物分子的开发与基于fret的HTS筛选的整合代表了一项独特的努力，通过实施药物发现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一个综合的视角，了解生物化学、现代遗传学和HTS之间的重要接口和协同作用。

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.