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遗传可编程模块 加州大学河滨分校CBET-0755775智力优势该合作项目的总体目标是开发一种遗传可编程模块，该模块易于适用于筛选各种蛋白酶的抑制剂。该方法是生成量子点（QD）修饰的蛋白酶特异性蛋白质模块，其可用作荧光共振能量转移（FRET）底物以探测蛋白酶活性。 这将导致新一代的遗传可编程蛋白质模块作为基于QD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QD的FRET底物，可以通过细胞穿透达特肽在细胞内传递。 该组合将使主要研究者提供一个快速和敏感的评估蛋白酶活性和相应的抑制剂的功效在HTS mathematics.Broader的影响：可调生物分子的发展与FRET为基础的HTS筛选的整合代表了一个独特的努力，扩大了蛋白质工程的基本发展与药物发现的实施。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HTS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.