Cationic Europium Complexes for Visualizing Fluctuations in Mitochondrial ATP Levels in Living Cells.
Cationic Europium Complexes for Visualizing Fluctuations in Mitochondrial ATP Levels in Living Cells.
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DOI：
10.1002/chem.201801008
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发表时间：
2018-07-25
期刊：
影响因子：
--
通讯作者：
Butler SJ
中科院分区：
文献类型：
--
作者：
Mailhot R;Traviss-Pollard T;Pal R;Butler SJ
由于缺乏能够快速、灵敏地显示细胞内三磷酸腺苷(ATP)浓度和分布的成像探针，阻碍了研究涉及三磷酸腺苷(ATP)的细胞代谢和酶过程的能力。我们在这里报道了第一个发光的稀土络合物的例子，它能够可视化活细胞线粒体中ATP浓度的变化。合成了四种阳离子Eu(III)配合物[Eu.1-4]+，考察了它们在生理pH水溶液中对核苷聚磷酸阴离子的结合能力。在悬臂上含有氢键供体基团的配合物[Eu.1]+和[Eu3]+对ATP、ADP和单磷酸物种表现出很好的区分能力。络合物[Eu]+表现出与ATP(logKa=5.8)的较强结合，提供了快速、长寿命的发光信号，使其能够在竞争激烈的水介质中检测到，其中包含生物相关浓度的镁、ADP、GTP、UTP和人血清白蛋白。该配合物在生理浓度范围内(1-5 mM)与三磷酸腺苷呈线性关系，并用于体外连续监测apyrase催化的三磷酸腺苷的水解反应。我们证明[EU.3]+可以有效地渗透哺乳动物(NIH-3T3)细胞，并选择性地定位于线粒体，从而允许实时观察在广谱激酶抑制剂星形孢子素治疗后线粒体ATP水平的升高，以及在葡萄糖饥饿条件下用氰化钾处理后线粒体ATP水平的下降。
The ability to study cellular metabolism and enzymatic processes involving adenosine triphosphate (ATP) is impeded by the lack of imaging probes capable of signalling the concentration and distribution of intracellular ATP rapidly, with high sensitivity. We report here the first example of a luminescent lanthanide complex capable of visualizing changes in the concentration of ATP in the mitochondria of living cells. Four cationic europium(III) complexes [Eu.1–4]+ have been synthesized and their binding capabilities towards nucleoside polyphosphate anions examined in aqueous solution at physiological pH. Complexes [Eu.1]+ and [Eu.3]+ bearing hydrogen bond donor groups in the pendant arms showed excellent discrimination between ATP, ADP and monophosphate species. Complex [Eu.3]+ showed relatively strong binding to ATP (logK a=5.8), providing a rapid, long‐lived luminescent signal that enabled its detection in a highly competitive aqueous medium containing biologically relevant concentrations of Mg2+, ADP, GTP, UTP and human serum albumin. This EuIII complex responds linearly to ATP within the physiological concentration range (1–5 mm), and was used to continuously monitor the apyrase‐catalyzed hydrolysis of ATP to ADP in vitro. We demonstrate that [Eu.3]+ can permeate mammalian (NIH‐3T3) cells efficiently and localize to the mitochondria selectively, permitting real‐time visualization of elevated mitochondrial ATP levels following treatment with a broad spectrum kinase inhibitor, staurosporine, as well as depleted ATP levels upon treatment with potassium cyanide under glucose starvation conditions.
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