A targetable fluorescent sensor reveals that copper-deficient SCO1 and SCO2 patient cells prioritize mitochondrial copper homeostasis.
A targetable fluorescent sensor reveals that copper-deficient SCO1 and SCO2 patient cells prioritize mitochondrial copper homeostasis.
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DOI：
10.1021/ja2004158
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发表时间：
2011-06-08
影响因子：
15
通讯作者：
Chang, Christopher J.
中科院分区：
文献类型：
--
作者：
Dodani, Sheel C.;Leary, Scot C.;Cobine, Paul A.;Winge, Dennis R.;Chang, Christopher J.
我们介绍了线粒体铜传感器-1 （Mito-CS1）的设计、合成、光谱学和生物学应用，这是一种用于成像活细胞中可交换线粒体铜池的新型靶向荧光传感器。Mito-CS1是一种双功能报告细胞，结合了Cu+响应荧光平台和线粒体靶向三苯磷片段，用于定位探针到该细胞器。Mito-CS1分子成像证实，这种新的化学工具可以检测HEK 293T模型细胞系和人成纤维细胞中不稳定线粒体Cu+的变化。此外，我们利用Mito-CS1对细胞色素c氧化酶1和2蛋白（SCO1和SCO2）合成突变患者的成纤维细胞进行了影像学和生化研究，这两种蛋白都是功能性活性细胞色素c氧化酶（COX）的组装和金属化所必需的。有趣的是，我们观察到，尽管这些线粒体金属伴侣的缺陷导致全细胞水平的铜缺乏，但与野生型对照相比，SCO1和SCO2患者成纤维细胞中的总铜和可交换线粒体Cu+池在很大程度上没有改变。我们的研究结果表明，即使在铜缺乏和线粒体金属伴侣功能障碍的情况下，细胞也能在线粒体中维持铜的稳态，这说明了调节铜在这种产能细胞器中的储存的重要性。
We present the design, synthesis, spectroscopy, and biological applications of Mitochondrial Coppersensor-1 (Mito-CS1), a new type of targetable fluorescent sensor for imaging exchangeable mitochondrial copper pools in living cells. Mito-CS1 is a bifunctional reporter that combines a Cu+-responsive fluorescent platform with a mitochondrial-targeting triphenylphosphonium moiety for localizing the probe to this organelle. Molecular imaging with Mito-CS1 establishes that this new chemical tool can detect changes in labile mitochondrial Cu+ in a model HEK 293T cell line as well as in human fibroblasts. Moreover, we utilized Mito-CS1 in a combined imaging and biochemical study in fibroblasts derived from patients with mutations in the two synthesis of cytochrome c oxidase 1 and 2 proteins (SCO1 and SCO2), each of which is required for assembly and metallation of functionally active cytochrome c oxidase (COX). Interestingly, we observe that although defects in these mitochondrial metallochaperones lead to a global copper deficiency at the whole cell level, total copper and exchangeable mitochondrial Cu+ pools in SCO1 and SCO2 patient fibroblasts are largely unaltered relative to wildtype controls. Our findings reveal that the cell maintains copper homeostasis in mitochondria even in situations of copper deficiency and mitochondrial metallochaperone malfunction, illustrating the importance of regulating copper stores in this energy-producing organelle.
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影响因子：
3
作者：
Banci, Lucia;Bertini, Ivano;Wang, Shenlin
通讯作者：
Wang, Shenlin
DOI：
10.1006/bbrc.1999.1073
发表时间：
1999-08-02
影响因子：
3.1
作者：
Camakaris, J;Voskoboinik, I;Mercer, JF
通讯作者：
Mercer, JF
影响因子：
4.8
作者：
Cobine, Paul A.;Pierrel, Fabien;Winge, Dennis R.
通讯作者：
Winge, Dennis R.
DOI：
10.1073/pnas.0800019105
发表时间：
2008-05-13
影响因子：
11.1
作者：
Banci, Lucia;Bertini, Ivano;Palumaa, Peep
通讯作者：
Palumaa, Peep
影响因子：
62.1
作者：
Atkinson, Aaron;Winge, Dennis R.
通讯作者：
Winge, Dennis R.