Establishment of Molecular Design Strategy To Obtain Activatable Fluorescent Probes for Carboxypeptidases
Establishment of Molecular Design Strategy To Obtain Activatable Fluorescent Probes for Carboxypeptidases
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DOI：
10.1021/jacs.7b11014
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发表时间：
2018-02-07
影响因子：
15
通讯作者：
Urano, Yasuteru
中科院分区：
文献类型：
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作者：
Kuriki, Yugo;Kamiya, Mako;Urano, Yasuteru
羧肽酶（Carboxypeptidases，CP）是一类水解酶，能从肽或蛋白质的C-末端切割一个或多个氨基酸。然而，监测国家协商进程活动的方法却很不完善。在这里，我们提出了第一个通用的设计策略，以获得可激活的荧光探针的CP通过利用分子内螺环化的罗丹明翻译的“脂肪族羧酰胺脂肪族羧酸酯”的结构转换催化的CP到动态荧光激活。基于这种新的策略，我们开发了羧肽酶A和B的探针。其中一种探针能够检测离体小鼠的胰液渗漏，表明其适用于胰瘘的术中诊断。这种设计策略应广泛适用于CP，以及其他以前不可靶向的酶，使荧光探针的发展，研究各种病理和生物过程。
Carboxypeptidases (CPs) are a family of hydrolases that cleave one or more amino acids from the C-terminal of peptides or proteins. However, methodology to monitor the activities of CPs is poorly developed. Here, we present the first versatile design strategy to obtain activatable fluorescent probes for CPs by utilizing intramolecular spirocyclization of rhodamine to translate the "aliphatic carboxamide to aliphatic carboxylate" structural conversion catalyzed by CPs into dynamic fluorescence activation. Based on this novel strategy, we developed probes for carboxypeptidases A and B. One of these probes was able to detect pancreatic juice leakage in mice ex vivo, suggesting that its suitability for intraoperative diagnosis of pancreatic fistula. This design strategy should be broadly applicable to CPs, as well as other previously untargetable enzymes, enabling development of fluorescent probes to study various pathological and biological processes.