Detection of Apoptosis Using Cyclic Luciferase in Living Mammals
Detection of Apoptosis Using Cyclic Luciferase in Living Mammals
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DOI：
10.1007/978-1-60327-321-3_9
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发表时间：
2009-01-01
期刊：
影响因子：
--
通讯作者：
Ozawa, Takeaki
中科院分区：
文献类型：
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作者：
Kanno, Akira;Umezawa, Yoshio;Ozawa, Takeaki
细胞程序性死亡即细胞凋亡是参与疾病发生发展的重要过程，由半胱氨酸蛋白酶（caspase）介导。活体中的半胱天冬酶活性及其与小化合物的调节对于筛选候选药物或病理因子具有极大的兴趣。我们描述了一种新的基因编码的生物发光指示剂的实时成像的半胱天冬酶-3在活细胞和动物的活动。该指示剂由工程萤火虫荧光素酶组成，其N-和C-末端与半胱天冬酶-3（Asp-Glu-Val-Asp）的底物序列连接。当激活的半胱天冬酶-3取代底物序列时，环化的荧光素酶恢复其活性。该指示剂提供了评价细胞毒性化合物或新型药理学化学品和细胞凋亡作用的一般方法。
Programmed cell death, apoptosis, is a crucial process involved in pathogenesis and progression of diseases, which is executed by cysteine aspartyl proteases (caspases). The caspase activities in living subjects and their regulation with small chemical compounds are of great interest for screening drug candidates or pathological agents. We describe a novel genetically encoded bioluminescent indicator for real-time imaging of caspase-3 activities in living cells and animals. The indicator is composed of an engineered firefly luciferase, of which the N- and C-terminal ends are linked with a substrate sequence of caspase-3 (Asp-Glu-Val-Asp). When activated caspase-3 digests the Substrate sequence, the cyclized luciferase recovers its activity. The indicator provides a general means of evaluating effects of cytotoxic Compounds or novel pharmacological chemicals and apoptosis.