一种鉴定蛋白酶体核心颗粒小分子刺激物的灵敏高通量筛选方法

Q3 Biochemistry, Genetics and Molecular Biology Current protocols in chemical biology Pub Date : 2018-10-04 DOI:10.1002/cpch.52

Rachel A. Coleman, Darci J. Trader

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引用次数: 4

摘要

荧光共振能量转移(FRET)技术是监测蛋白质相互作用和蛋白酶活性的有用工具。我们最近报道了一项生化试验，利用FRET报告肽来监测蛋白酶体的20S催化颗粒(20S CP)的活性。该试验专门设计用于提高识别20S CP刺激因子的敏感性，这可能具有治疗蛋白质蓄积性疾病的治疗潜力。本文描述的方案详细介绍了合成FRET报告肽和使用20S CP进行FRET测定的必要步骤。©2018 by John Wiley &儿子,Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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A Sensitive High-Throughput Screening Method for Identifying Small Molecule Stimulators of the Core Particle of the Proteasome

Fluorescence resonance energy transfer (FRET) technology is a useful tool to monitor protein interactions as well as protease activity. We have recently reported a biochemical assay utilizing a FRET reporter peptide to monitor the activity of the 20S catalytic particle (20S CP) of the proteasome. This assay is designed specifically to have increased sensitivity to identify stimulators of the 20S CP, which may hold therapeutic potential to treat protein-accumulation diseases. The protocol described here details the necessary steps in synthesizing the FRET reporter peptide and performing the FRET assay with the 20S CP. © 2018 by John Wiley & Sons, Inc.

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Current protocols in chemical biology Biochemistry, Genetics and Molecular Biology-Biophysics

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