An Endoperoxide Reactivity-Based FRET Probe for Ratiometric Fluorescence Imaging of Labile Iron Pools in Living Cells.
An Endoperoxide Reactivity-Based FRET Probe for Ratiometric Fluorescence Imaging of Labile Iron Pools in Living Cells.
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DOI：
10.1021/jacs.6b08016
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发表时间：
2016-11-02
影响因子：
15
通讯作者：
Chang CJ
中科院分区：
文献类型：
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作者：
Aron AT;Loehr MO;Bogena J;Chang CJ
铁对于维持生命是必不可少的，因为它在多种氧化状态之间循环的能力对于催化生物系统中的化学转化至关重要。然而，如果没有适当的调控，这种同样的氧化还原能力会引发氧化应激事件，导致衰老，以及从癌症到心血管和神经退行性疾病等各种疾病。尽管生物铁很重要，但由于铁具有很强的荧光猝灭能力，监测生物铁与细胞配体−不稳定的铁池−结合以产生保存空间和时间信息的反应的方法仍然有限。我们报道了FRET铁探针1(FIP-1)的设计、合成和生物学评价，FIP-1是一种基于反应性的探针，可以对生命系统中不稳定的铁池进行比率荧光成像。受到抗疟疾天然产物和相关治疗药物的启发，FIP-1通过Fe(II)可切割的过氧化内桥将两个荧光素(荧光素和Cy3)连接起来，其中Fe(II)引发的过氧化氢切割通过将这两种染料裂解成单独的片段，减少了从荧光素供体到Cy3受体的荧光共振能量转移(FRET)。FIP-1对水缓冲溶液中的Fe(II)具有选择性，对竞争的金属离子具有选择性，并能够检测在铁补充和/或铁耗竭情况下活细胞内不稳定铁库的变化。此外，将FIP-1应用于铁下垂模型，揭示了在这种形式的细胞死亡过程中不稳定铁库的变化，为研究生命系统中的铁信号提供了一个起点。
Iron is essential for sustaining life, as its ability to cycle between multiple oxidation states is critical for catalyzing chemical transformations in biological systems. However, without proper regulation, this same redox capacity can trigger oxidative stress events that contribute to aging along with diseases ranging from cancer to cardiovascular and neurodegenerative disorders. Despite its importance, methods for monitoring biological iron bound weakly to cellular ligands−the labile iron pool−to generate a response that preserves spatial and temporal information remain limited, owing to the potent fluorescence quenching ability of iron. We report the design, synthesis, and biological evaluation of FRET Iron Probe 1 (FIP-1), a reactivity-based probe that enables ratiometric fluorescence imaging of labile iron pools in living systems. Inspired by antimalarial natural products and related therapeutics, FIP-1 links two fluorophores (fluorescein and Cy3) through an Fe(II)-cleavable endoperoxide bridge, where Fe(II)-triggered peroxide cleavage leads to a decrease in fluorescence resonance energy transfer (FRET) from the fluorescein donor to Cy3 acceptor by splitting these two dyes into separate fragments. FIP-1 responds to Fe(II) in aqueous buffer with selectivity over competing metal ions and is capable of detecting changes in labile iron pools within living cells with iron supplementation and/or depletion. Moreover, application of FIP-1 to a model of ferroptosis reveals a change in labile iron pools during this form of cell death, providing a starting point to study iron signaling in living systems.
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影响因子：
46.2
作者：
Cho DG;Sessler JL
通讯作者：
Sessler JL
影响因子：
64.5
作者：
Dixon SJ;Lemberg KM;Lamprecht MR;Skouta R;Zaitsev EM;Gleason CE;Patel DN;Bauer AJ;Cantley AM;Yang WS;Morrison B 3rd;Stockwell BR
通讯作者：
Stockwell BR
影响因子：
2.9
作者：
Epsztejn, S;Kakhlon, O;Cabantchik, ZI
通讯作者：
Cabantchik, ZI
DOI：
10.1073/pnas.1312782110
发表时间：
2013-11-05
影响因子：
11.1
作者：
Deu, Edgar;Chen, Ingrid T.;Bogyo, Matthew
通讯作者：
Bogyo, Matthew
影响因子：
21.8
作者：
通讯作者：
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