Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues,Chemical Science

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Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues
Chemical Science ( IF 7.6 ) Pub Date : 2017-09-11 00:00:00 , DOI: 10.1039/c7sc02805a
Michael W. Beck _{1,

2,

3,

4} , Rahul S. Kathayat _{1,

2,

3,

4} , Candace M. Cham _{2,

3,

4,

5} , Eugene B. Chang _{2,

3,

4,

5} , Bryan C. Dickinson _{1,

2,

3,

4}

Affiliation

The reversible modification of cysteine residues through thioester formation with palmitate (protein S-palmitoylation) is a prevalent chemical modification that regulates the function, localization, and stability of many proteins. Current methods for monitoring the “erasers” of S-palmitoylation, acyl-protein thioesterases (APTs), rely on destructive proteomic methods or “turn-on” probes, precluding deployment in heterogeneous samples such as primary tissues. To address these challenges, we present the design, synthesis, and biological evaluation of Ratiometric Depalmitoylation Probes (RDPs). RDPs respond to APTs with a robust ratiometric change in fluorescent signal both in vitro and in live cells. Moreover, RDPs can monitor endogenous APT activities in heterogeneous primary human tissues such as colon organoids, presaging the utility of these molecules in uncovering novel roles for APTs in metabolic regulation.

中文翻译：

基于迈克尔加成的探针，用于在活细胞和组织中对蛋白质S-去棕榈糖进行比例荧光成像

通过与棕榈酸酯形成硫酯形成的半胱氨酸残基的可逆修饰（蛋白质S-棕榈酰化）是一种常见的化学修饰，它调节许多蛋白质的功能，定位和稳定性。目前用于监测S-棕榈酰化的“清除剂” ，酰基蛋白硫酯酶（APT）的方法依赖于破坏性的蛋白质组学方法或“开启”探针，从而阻止了在异质样品（例如原发组织）中的部署。为了解决这些挑战，我们介绍了比例去棕榈酸酯化探针（RDP）的设计，合成和生物学评估。RDP对APT的反应均在体外产生强烈的荧光信号比率变化和活细胞中。此外，RDP可以监测异种主要人类组织（例如结肠类器官）中的内源性APT活性，从而预示了这些分子在揭示APT在代谢调节中的新作用中的效用。

更新日期：2017-09-22

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