4 April 2005 Dynamic detection of caspase-3 activation during photosensitization by fluorescence resonance energy transfer
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Abstract
Apoptosis is one of the important modes in PDT-induced cell death. Activation of caspase-3 is considered to be the final step in many apoptosis pathways. In this study, we used SCAT3, a fluorescence resonance energy transfer (FRET) probe containing caspase-3 substrate, to study the dynamics of caspase-3 activation in living ASTC-a-1 cells expressing stably SCAT3. The FRET analysis results indicated that caspase-3 activation in response to tumor necrosis factor-α or PDT resulted in cleavage of the linker peptide and subsequent disruption of the FRET signal. The SCAT3 was cleaved immediately after PDT treatment, but that for TNF-a treatment was delayed two hours. Our experimental results suggested that the different apoptotic pathways induced by TNF-α or PDT caused different cleavage kinetics of SCAT3. This study shows that FRET technique based on GFPs could be used to study the mechanism of PDT-induced apoptosis in living cells.
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Yunxia Wu, Yunxia Wu, Da Xing, Da Xing, Qun Chen, Qun Chen, Tongsheng Chen, Tongsheng Chen, Yonghong Tang, Yonghong Tang, Qingling Wan, Qingling Wan, } "Dynamic detection of caspase-3 activation during photosensitization by fluorescence resonance energy transfer", Proc. SPIE 5704, Genetically Engineered and Optical Probes for Biomedical Applications III, (4 April 2005); doi: 10.1117/12.588963; https://doi.org/10.1117/12.588963
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