Real-time detection of caspase-2 activation in a single living HeLa cell during cisplatin-induced apoptosis

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....

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Bibliographic Details
Published in:Journal of Biomedical Optics 2006-03, Vol.11 (2), p.024011-024016
Main Authors: Lin, Juqiang, Zhang, Zhihong, Yang, Jie, Zeng, Shaoqun, Liu, Bi-Feng, Luo, Qingming
Format: Article
Language:English
Subjects:
Antineoplastic Agents - administration & dosage
Apoptosis - drug effects
Caspase 2
Caspases - drug effects
Caspases - metabolism
cisplatin
Cisplatin - administration & dosage
Computer Systems
cyan fluorescence protein
Enzyme Activation - drug effects
fluorescence resonance energy transfer
Fluorescence Resonance Energy Transfer - methods
Green Fluorescent Proteins
HeLa Cells
Humans
Luminescent Proteins
red fluorescence protein from
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Summary: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.
ISSN:1083-3668
1560-2281
DOI:10.1117/1.2187013