A highly sensitive and selective two-photon fluorescent probe for real-time sensing of cytochrome P450 1A1 in living systems†,Materials Chemistry Frontiers

当前位置： X-MOL 学术 › Mater. Chem. Front. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

A highly sensitive and selective two-photon fluorescent probe for real-time sensing of cytochrome P450 1A1 in living systems†
Materials Chemistry Frontiers ( IF 6.0 ) Pub Date : 2018-09-07 00:00:00 , DOI: 10.1039/c8qm00372f
Jing Ning _{1,

2,

3,

4,

5} , Zhenhao Tian _{6,

7,

8,

9} , Bo Wang _{1,

2,

3,

4,

5} , Guangbo Ge _{10,

11,

12} , Yue An _{1,

2,

3,

4,

5} , Jie Hou _{1,

2,

3,

4,

5} , Chao Wang _{1,

2,

3,

4,

5} , Xinyu Zhao _{1,

2,

3,

4,

5} , Yannan Li _{1,

2,

3,

4,

5} , Xiangge Tian _{1,

2,

3,

4,

5} , Zhenlong Yu _{1,

2,

3,

4,

5} , Xiaokui Huo _{1,

2,

3,

4,

5} , Chengpeng Sun _{1,

2,

3,

4,

5} , Lei Feng _{1,

2,

3,

4,

5} , Jingnan Cui _{6,

7,

8,

9} , Xiaochi Ma _{1,

2,

3,

4,

5}

Affiliation

Cytochrome P450 1A1 (CYP1A1), a heme-containing monooxygenase, is of particular importance for human health because of its vital role in the metabolic activation of pro-carcinogenic compounds to their ultimate carcinogens. However, CYP1A1 protein levels are extraordinarily low in normal and cancer tissues. Thus, a practical method for ultra-sensitive and real-time monitoring of CYP1A1 activity in complex biological systems is highly sought after. In the present study, we developed a highly specific and sensitive two-photon fluorescent probe for monitoring CYP1A1 activity on the basis of the substrate preferences of this key enzyme. A panel of O-alkylated derivatives was designed and synthesized using HBN as the basic fluorophore. After screening and optimization, the derivative iPrBN was selected for further study as it displayed excellent specificity, high sensitivity and fast turn-on response to CYP1A1 relative to other human CYP isoforms. The detection limit of iPrBN for CYP1A1 was 0.036 nM, suggesting that it would be sensitive and versatile enough to detect endogenous CYP1A1 activity. Indeed, we successfully applied iPrBN to the real-time monitoring of CYP1A1 activity in human cancer cell lines and performed high-throughput screening of CYP1A1 modulators. iPrBN was also applied for the first time to the two-photon imaging of intracellular CYP1A1 in living cancer tissues and zebrafish, and our results showed that iPrBN exhibited high imaging resolution and fast response towards CYP1A1. These findings suggest that this probe is capable of accurately sensing CYP1A1 activity in complex biological systems, which will facilitate further investigations of CYP1A1-associated physiological and pathological processes.

中文翻译：

一种高灵敏度和选择性的双光子荧光探针，用于实时检测生命系统中的细胞色素P450 1A1 †

细胞色素P450 1A1（CYP1A1）是一种含血红素的单加氧酶，对人类健康尤为重要，因为它在致癌性化合物代谢为最终致癌物的新陈代谢中起着至关重要的作用。但是，正常组织和癌症组织中的CYP1A1蛋白水平异常低。因此，人们迫切需要一种实用的方法来对复杂生物系统中的CYP1A1活性进行超灵敏的实时监控。在本研究中，我们基于该关键酶的底物偏好，开发了一种高特异性和灵敏的双光子荧光探针，用于监测CYP1A1的活性。使用HBN作为碱性荧光团设计和合成了一组O烷基化衍生物。经过筛选和优化后，导数iPrBN被选择作进一步研究，因为它相对于其他人CYP同工型显示出优异的特异性，高灵敏度和对CYP1A1的快速开启反应。CYP1A1的iPrBN的检出限为0.036 nM，这表明它对检测内源性CYP1A1活性具有足够的灵敏性和通用性。确实，我们成功地将iPrBN应用于人癌细胞系中CYP1A1活性的实时监测，并进行了CYP1A1调节剂的高通量筛选。iPrBN也首次应用于活体癌组织和斑马鱼细胞内CYP1A1的双光子成像，我们的结果表明iPrBN表现出高成像分辨率和对CYP1A1的快速响应。这些发现表明，该探针能够在复杂的生物系统中准确检测CYP1A1的活性，这将有助于进一步研究CYP1A1相关的生理和病理过程。

更新日期：2018-09-07

点击分享查看原文

点击收藏

阅读更多本刊最新论文