Reaction-based fluorescent sensor for investigating mobile Zn2+ in mitochondria of healthy versus cancerous prostate cells
Reaction-based fluorescent sensor for investigating mobile Zn2+ in mitochondria of healthy versus cancerous prostate cells
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DOI：
10.1073/pnas.1310583110
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发表时间：
2014-01-07
影响因子：
11.1
通讯作者：
Radford, Robert J.
中科院分区：
文献类型：
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作者：
Chyan, Wen;Zhang, Daniel Y.;Radford, Robert J.
可螯合的、移动的形式的二价锌Zn（II）在哺乳动物生物学中起着重要的信号传导作用。锌转运蛋白的复杂网络已经进化到在亚细胞水平上控制锌的浓度和分布。了解行动的移动的锌需要的工具，可以检测锌（II）浓度的变化在离散的细胞区域。我们在这里提出了一个锌响应，反应为基础的，有针对性的探针的基础上的双乙酰化形式的Zinpyr-1。该化合物，（6-氨基乙基）三苯基磷锌吡-1二乙酸盐（DA-ZP 1-TPP），在无金属状态下基本上是无荧光的;然而，暴露于Zn（II）触发金属介导的乙酰基水解，以提供大的，快速的，锌诱导的荧光响应。DA-ZP 1-TPP在2小时内对细胞内酯酶不敏感，并且不受质子诱导的开启的影响。DA-ZP 1-TPP的实际效用通过实验证明，该实验揭示，与健康的上皮前列腺细胞相反，致瘤细胞不能在其线粒体内积累移动的锌。
Chelatable, mobile forms of divalent zinc, Zn(II), play essential signaling roles in mammalian biology. A complex network of zinc import and transport proteins has evolved to control zinc concentration and distribution on a subcellular level. Understanding the action of mobile zinc requires tools that can detect changes in Zn(II) concentrations at discrete cellular locales. We present here a zinc-responsive, reaction-based, targetable probe based on the diacetyled form of Zinpyr-1. The compound, (6-amidoethyl)triphenylphosphonium Zinpyr-1 diacetate (DA-ZP1-TPP), is essentially nonfluorescent in the metal-free state; however, exposure to Zn (II) triggers metal-mediated hydrolysis of the acetyl groups to afford a large, rapid, and zinc-induced fluorescence response. DA-ZP1-TPP is insensitive to intracellular esterases over a 2-h period and is impervious to proton-induced turn-on. A TPP unit is appended for targeting mitochondria, as demonstrated by live cell fluorescence imaging studies. The practical utility of DA-ZP1-TPP is demonstrated by experiments revealing that, in contrast to healthy epithelial prostate cells, tumorigenic cells are unable to accumulate mobile zinc within their mitochondria.