Lin Juqiang, Zhang Zhihong, Yang Jie, Zeng Shaoqun, Liu Bi-Feng, Luo Qingming
Huazhong University of Science and Technology, Key Laboratory of Biomedical Photonics of Ministry of Education-Hubei, Bioinformatics and Molecular Imaging Key Laboratory, Wuhan 430074 China.
J Biomed Opt. 2006 Mar-Apr;11(2):024011. doi: 10.1117/1.2187013.
Caspase-2 is important for the mitochondrial apoptotic pathway, however, the mechanism by which caspase-2 executes apoptosis remains obscure. We carry out the first measurements of the dynamics of caspase-2 activation in a single living cell by a FRET (fluorescence resonance energy transfer) probe. Two FRET probes are constructed that each encoded a CRS (caspase-2 or caspase-3 recognition site) fused with a cyan fluorescent protein (CFP) and a red fluorescent protein (DsRed) (CFP-CRS-DsRed). Using these probes, we found that during cisplatin-induced apoptosis, caspase-2 activation occurred more slowly than did activation of caspase-3; additionally, caspase-2 activation was initiated much earlier than that of caspase-3.
半胱天冬酶-2对于线粒体凋亡途径很重要，然而，半胱天冬酶-2执行凋亡的机制仍不清楚。我们通过荧光共振能量转移（FRET）探针首次测量了单个活细胞中半胱天冬酶-2激活的动力学。构建了两种FRET探针，每种探针都编码一个与青色荧光蛋白（CFP）和红色荧光蛋白（DsRed）融合的半胱天冬酶-2或半胱天冬酶-3识别位点（CFP-CRS-DsRed）。使用这些探针，我们发现在顺铂诱导的凋亡过程中，半胱天冬酶-2的激活比半胱天冬酶-3的激活发生得更慢；此外，半胱天冬酶-2的激活比半胱天冬酶-3的激活起始得更早。
