基于熄灭荧光的高保真分析法检测细胞蛋白稳态的干扰

IF 3.8 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY ACS Bio & Med Chem Au Pub Date : 2024-01-19 DOI:10.1021/acsbiomedchemau.3c00012
Conner Hoelzel, Yulong Bai, Mengdie Wang, Yu Liu* and Xin Zhang*, 
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引用次数: 0

摘要

神经退行性疾病久治不愈,因此有必要开发监测蛋白质稳态(proteostasis)的新策略。我们实验室之前的工作重点是开发荧光策略,以观察蛋白稳态压力的发生和发展。这些工作利用溶解变色和粘度敏感荧光团来感知蛋白质的折叠状态,从而通过增加聚集时的发射强度来筛选应激源。在这项工作中,我们提出了一种新颖、高保真的检测方法,用于检测细胞蛋白稳态的扰动,荧光强度会随着蛋白稳态应激的发生而降低。利用含氟羟甲基硅-罗丹明探针来区分蛋白质折叠状态,我们在活细胞中证明了这一技术的有效性,在未受压和受压条件下荧光强度相差两倍。
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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High-Fidelity Assay Based on Turn-Off Fluorescence to Detect the Perturbations of Cellular Proteostasis

The persistence of neurodegenerative diseases has necessitated the development of new strategies to monitor protein homeostasis (proteostasis). Previous efforts in our laboratory have focused on the development of fluorogenic strategies to observe the onset and progression of proteostatic stress. These works utilized solvatochromic and viscosity sensitive fluorophores to sense protein folded states, enabling stressor screening with an increase in the emission intensity upon aggregation. In this work, we present a novel, high-fidelity assay to detect perturbations of cellular proteostasis, where the fluorescence intensity decreases with the onset of proteostatic stress. Utilizing a fluorogenic, hydroxymethyl silicon-rhodamine probe to differentiate between protein folded states, we establish the validity of this technology in living cells by demonstrating a two-fold difference in fluorescence intensity between unstressed and stressed conditions.

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来源期刊
ACS Bio & Med Chem Au
ACS Bio & Med Chem Au 药物、生物、化学-
CiteScore
4.10
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0.00%
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0
期刊介绍: ACS Bio & Med Chem Au is a broad scope open access journal which publishes short letters comprehensive articles reviews and perspectives in all aspects of biological and medicinal chemistry. Studies providing fundamental insights or describing novel syntheses as well as clinical or other applications-based work are welcomed.This broad scope includes experimental and theoretical studies on the chemical physical mechanistic and/or structural basis of biological or cell function in all domains of life. It encompasses the fields of chemical biology synthetic biology disease biology cell biology agriculture and food natural products research nucleic acid biology neuroscience structural biology and biophysics.The journal publishes studies that pertain to a broad range of medicinal chemistry including compound design and optimization biological evaluation molecular mechanistic understanding of drug delivery and drug delivery systems imaging agents and pharmacology and translational science of both small and large bioactive molecules. Novel computational cheminformatics and structural studies for the identification (or structure-activity relationship analysis) of bioactive molecules ligands and their targets are also welcome. The journal will consider computational studies applying established computational methods but only in combination with novel and original experimental data (e.g. in cases where new compounds have been designed and tested).Also included in the scope of the journal are articles relating to infectious diseases research on pathogens host-pathogen interactions therapeutics diagnostics vaccines drug-delivery systems and other biomedical technology development pertaining to infectious diseases.
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