Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues.
Michael addition-based probes for ratiometric fluorescence imaging of protein S-depalmitoylases in live cells and tissues.
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DOI：
10.1039/c7sc02805a
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发表时间：
2017-11-01
期刊：
影响因子：
8.4
通讯作者：
Dickinson BC
中科院分区：
文献类型：
--
作者：
Beck MW;Kathayat RS;Cham CM;Chang EB;Dickinson BC
半胱氨酸棕榈酰化“橡皮擦”的比例荧光探针允许活细胞和组织的内源性酶活性成像。半胱氨酸残基通过与棕榈酸酯形成硫酯（蛋白质S-棕榈酰化）的可逆修饰是一种普遍的化学修饰，其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“擦除器”、酰基蛋白硫酯酶（APT）的方法依赖于破坏性蛋白质组学方法或“开启”探针，从而排除了在异质样品如原代组织中的部署。为了解决这些挑战，我们提出了设计，合成和生物评价的比率脱棕榈酰化探针（RDPs）。RDP对APT的响应在体外和活细胞中均具有强的荧光信号比率变化。此外，RDP可以监测异质性原发性人体组织（如结肠类器官）中的内源性APT活性，预示着这些分子在揭示APT在代谢调节中的新作用方面的效用。
Ratiometric fluorescent probes for cysteine palmitoylation “erasers” permit live cell and tissue imaging of endogenous enzyme activities. 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.
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DOI：
10.1523/jneurosci.2171-08.2008
发表时间：
2008-09-03
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Ponimaskin E;Dityateva G;Ruonala MO;Fukata M;Fukata Y;Kobe F;Wouters FS;Delling M;Bredt DS;Schachner M;Dityatev A
通讯作者：
Dityatev A
影响因子：
14.8
作者：
Dekker, Frank J.;Rocks, Oliver;Waldmann, Herbert
通讯作者：
Waldmann, Herbert
影响因子：
9
作者：
通讯作者：
--
半胱氨酸二光二酰化的荧光探针揭示了动态的APT信号传导。
影响因子：
14.8
作者：
Kathayat RS;Elvira PD;Dickinson BC
通讯作者：
Dickinson BC
影响因子：
3.9
作者：
Kim, Gun-Joong;Yoon, Doo-Ha;Kim, Hae-Jo
通讯作者：
Kim, Hae-Jo