Multiplex imaging of an intracellular proteolytic cascade by using a broad-spectrum nanoquencher.
Multiplex imaging of an intracellular proteolytic cascade by using a broad-spectrum nanoquencher.
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DOI：
10.1002/anie.201107795
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发表时间：
2012-02-13
影响因子：
16.6
通讯作者：
Chen, Xiaoyuan
中科院分区：
文献类型：
--
作者：
Huang, Xinglu;Swierczewska, Magdalena;Choi, Ki Young;Zhu, Lei;Bhirde, Ashwinkumar;Park, Jinwoo;Kim, Kwangmeyung;Xie, Jin;Niu, Gang;Lee, Kang Choon;Lee, Seulki;Chen, Xiaoyuan
蛋白水解酶是一种极其重要的信号分子，对特定蛋白水解酶活性的破坏最终会导致严重的疾病，包括心血管疾病、骨质疏松症、炎症性疾病、神经退行性疾病和癌症。[1]因此，确定特定蛋白水解酶在特定生物过程中的作用对于促进疾病预防的新方法至关重要。[2]一个能够直接和同时监测生物体内多种蛋白分解活性的系统对于阐明复杂的蛋白水解酶信号通路以及测试针对蛋白水解酶的药物的有效性具有巨大的潜力。到目前为止，已经开发了一系列基于荧光共振能量转移(FRET)的荧光肽，这些荧光肽由底物和染料/猝灭剂对结合组成，以监测蛋白水解性。[3]然而，传统的荧光肽在检测由多种蛋白水解酶诱导的蛋白水解级联中的蛋白酶信号方面作用有限，例如，凝血级联。此外，使用荧光肽来产生多颜色的蛋白酶图像将需要多步合成和独特的染料/猝灭剂对选择。使用广谱猝灭剂可能是合适的，但这种分子不能完全猝灭所有可用的染料，而且合成也不方便。此外，为了针对细胞内的蛋白酶，传感器需要进一步用穿透性多肽或转染剂进行修饰以进入细胞。[6]可激活传感器领域的最新进展使体外和体内各种蛋白酶活性的实时成像成为可能。[7]然而，没有一个传感器被用于活细胞中的细胞内蛋白酶的多路成像。工程化的荧光蛋白FRET对和伴随着蛋白酶底物的发光蛋白也被开发用于实时细胞成像，但这些策略需要转基因细胞。
Proteases are known as extremely important signaling molecules and the deregulation of specific protease activities can ultimately lead to severe pathologies, which include cardiovascular disease, osteoporosis, inflammatory disease, neurodegenerative disorders, and cancer.[1] Therefore, identifying the role of a specific protease in a given biological process is crucial to promote new approaches for the prevention of diseases.[2] A system that enables the direct and simultaneous monitoring of multiple proteolytic activities in living organisms has great potential for elucidating complex protease signaling pathways, as well as testing the efficacy of drugs that target proteases.To date, a range of fluorescence resonance energy transfer (FRET) based fluorogenic peptides that are comprised of a substrate combined with a dye/quencher pair have been developed to monitor proteolytic activities.[3] However, the conventional fluorogenic peptides are of limited use for examining protease signaling in a proteolytic cascade that is induced by multiple proteases, for example, the blood coagulation cascade.,[4] Furthermore, using a fluorogenic peptide for producing multicolored images of proteases would require a multistep synthesis and distinctive dye/quencher pair selections. The use of broad-spectrum quenchers [5] may be appropriate, but such molecules do not completely quench all of the available dyes, and are also inconvenient to synthesize. Moreover, to target intracellular proteases, sensors need to be further modified with cell-penetrating peptides or transfecting agents to be taken up into cells.[6] Recent advances in the field of activable sensors have enabled real-time imaging of various protease activities in vitro and in vivo.[7] However, no single sensor has been used for multiplexed imaging of intracellular proteases in living cells. Engineered fluorescent protein FRET pairs and luminescent proteins that are accompanied by a protease substrate have also been developed for real-time cellular imaging, but these strategies require genetically modified cells.[8]
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影响因子：
2.8
作者：
Cudic M;Fields GB
通讯作者：
Fields GB
影响因子：
2.9
作者：
Peng, Xinzhan;Chen, Huaxian;Olive, D. Michael
通讯作者：
Olive, D. Michael
影响因子：
4.7
作者：
Lee, Seulki;Choi, Ki Young;Choi, Kuiwon
通讯作者：
Choi, Kuiwon
DOI：
10.1073/pnas.0907367106
发表时间：
2009-10-20
影响因子：
11.1
作者：
Jun, Young-Wook;Sheikholeslami, Sassan;Alivisatos, A. Paul
通讯作者：
Alivisatos, A. Paul
DOI：
10.1002/anie.201007805
发表时间：
2011-05-23
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Dai N;Guo J;Teo YN;Kool ET
通讯作者：
Kool ET