Non-peptide-based fluorogenic small-molecule probe for elastase.
Non-peptide-based fluorogenic small-molecule probe for elastase.
复制标题
用于弹性蛋白酶的非肽荧光小分子探针。
DOI：
10.1021/ac402097g
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发表时间：
2013-11
影响因子：
7.4
通讯作者：
Yang, Guang-Fu
中科院分区：
文献类型：
--
作者：
Sun, Qi;Li, Jun;Liu, Wan-Nian;Dong, Qing-Jian;Yang, Wen-Chao;Yang, Guang-Fu
人中性粒细胞弹性蛋白酶(HNE)几十年来一直被认为是发现抗炎药物的潜在治疗靶点。然而，在测定HNE活性方面进展甚微，尤其是在测定HNE活性中起关键作用的合成底物上。在这里，一种小分子化合物2，2，3，3，3-pentafluoro-N-(2-oxo-4-(trifluoromethyl)-2H-chromen-7-yl)-propanamide(化合物4)被成功地设计为有史以来第一个非基于多肽的HNE荧光底物。建立了一种基于4的“开启”荧光法，用于HNE活性的快速测定和抑制动力学研究。最重要的是，探针4对7种被测试的水解酶或蛋白质中的HNE显示出高度的特异性反应，并可直接用于检测慢性阻塞性肺疾病(COPD)患者与健康对照组相比血清中HNE活性的升高。这种特异且经济有效的探针将有助于未来高通量发现HNE抑制剂和弹性蛋白酶相关疾病的临床诊断。
Human neutrophil elastase (HNE) has been identified as a potential therapeutic target for the discovery of anti-inflammatory drugs for decades. However, little progress has been made on assays measuring the activity of HNE, especially on synthetic substrates which play essential role in determination of HNE activity. Herein, a small-molecule compound, 2,2,3,3,3-pentafluoro-N-(2-oxo-4-(trifluoromethyl)-2H-chromen-7-yl)-propanamide (compound 4), has been successfully designed as the first ever non-peptide-based fluorogenic substrate for HNE. A "turn-on" fluorometric assay based on 4 has been successfully developed for rapid determination of HNE activity and the inhibitory kinetic study. Most importantly, the probe 4 shows highly specific response for HNE among seven tested hydrolases or proteins and can be directly used to detect the elevated HNE activity in the serum of chronic obstructive pulmonary disease (COPD) patients compared to that of healthy controls. This specific and cost-effective probe will facilitate future high-throughput discovery of HNE inhibitors and clinical diagnosis of elastase-related diseases.
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DOI：
10.1016/s0021-9258(18)50690-6
发表时间：
1979-05
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
K. Nakajima;J. Powers;B. Ashe;M. Zimmerman
通讯作者：
K. Nakajima;J. Powers;B. Ashe;M. Zimmerman
影响因子：
6.1
作者：
B. Zorn;I. Virant-Klun;H. Meden‐Vrtovec
通讯作者：
B. Zorn;I. Virant-Klun;H. Meden‐Vrtovec
影响因子：
15
作者：
Ros-Lis, JV;García, B;Valldecabres, MC
通讯作者：
Valldecabres, MC
影响因子：
1.6
作者：
E. Masler
通讯作者：
E. Masler
影响因子：
11.2
作者：
Mittendorf EA;Alatrash G;Qiao N;Wu Y;Sukhumalchandra P;St John LS;Philips AV;Xiao H;Zhang M;Ruisaard K;Clise-Dwyer K;Lu S;Molldrem JJ
通讯作者：
Molldrem JJ