利用荧光能量转移检测程序性细胞死亡
Detection of programmed cell death using fluorescence energy transfer.
作者信息
Xu X, Gerard A L, Huang B C, Anderson D C, Payan D G, Luo Y
机构信息
Rigel, Inc., 772 Lucerne Drive, Sunnyvale, CA 94086, USA.
出版信息
Nucleic Acids Res. 1998 Apr 15;26(8):2034-5. doi: 10.1093/nar/26.8.2034.
Abstract
Fluorescence energy transfer (FRET) can be generated when green fluorescent protein (GFP) and blue fluorescent protein (BFP) are covalently linked together by a short peptide. Cleavage of this linkage by protease completely eliminates FRET effect. Caspase-3 (CPP32) is an important cellular protease activated during programmed cell death. An 18 amino acid peptide containing CPP32 recognition sequence, DEVD, was used to link GFP and BFP together. CPP32 activation can be monitored by FRET assay during the apoptosis process.
摘要
当绿色荧光蛋白（GFP）和蓝色荧光蛋白（BFP）通过短肽共价连接在一起时，可产生荧光能量转移（FRET）。蛋白酶对这种连接的切割会完全消除FRET效应。半胱天冬酶-3（CPP32）是一种在程序性细胞死亡过程中被激活的重要细胞蛋白酶。一种含有CPP32识别序列DEVD的18个氨基酸的肽被用于将GFP和BFP连接在一起。在细胞凋亡过程中，可通过FRET测定法监测CPP32的激活情况。
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