| Measuring dynamics of caspase-8 activation in a single living HeLa cell during TNFalpha-induced apoptosis. | |
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MedLine Citation:
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PMID: 12711301 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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In this study, we reported the first measurement of the dynamics of activation of caspase-8 in a single living cell. This measurement was conducted using a specially developed molecular sensor based on the FRET (fluorescence resonance energy transfer) technique. This sensor was constructed by fusing a CFP (cyan fluorescent protein) and a YFP (yellow fluorescent protein) with a linker containing a tandem caspase-8-specific cleavage site. The change of the FRET ratio upon cleavage was larger than 4-fold. Using this sensor, we found that during TNFalpha-induced apoptosis, the activation of caspase-8 was a slower process than that of caspase-3, and it was initiated much earlier than the caspase-3 activation. Inhibition of caspase-9 delayed the full activation of caspase-3 but did not affect the dynamics of caspase-8. Results of these single-cell measurements suggested that caspase-3 was activated by caspase-8 through two parallel pathways during TNFalpha-induced apoptosis in HeLa cells. |
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Authors:
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Kathy Q Luo; Vivian C Yu; Yongmei Pu; Donald C Chang |
Publication Detail:
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Type: Evaluation Studies; Journal Article |
Journal Detail:
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Title: Biochemical and biophysical research communications Volume: 304 ISSN: 0006-291X ISO Abbreviation: Biochem. Biophys. Res. Commun. Publication Date: 2003 May |
Date Detail:
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Created Date: 2003-04-24 Completed Date: 2003-06-18 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0372516 Medline TA: Biochem Biophys Res Commun Country: United States |
Other Details:
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Languages: eng Pagination: 217-22 Citation Subset: IM |
Affiliation:
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Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, China. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Apoptosis* Bacterial Proteins / chemistry, genetics Caspase 3 Caspase 8 Caspase 9 Caspases / metabolism* Enzyme Activation Fluorescence Resonance Energy Transfer* Fluorescent Dyes / chemistry Green Fluorescent Proteins Hela Cells Humans Luminescent Proteins / chemistry, genetics Microscopy, Fluorescence Models, Biological Molecular Sequence Data Recombinant Fusion Proteins / analysis Signal Transduction Tumor Necrosis Factor-alpha / pharmacology |
| Chemical | |
Reg. No./Substance:
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0/Bacterial Proteins; 0/Fluorescent Dyes; 0/Luminescent Proteins; 0/Recombinant Fusion Proteins; 0/Tumor Necrosis Factor-alpha; 0/yellow fluorescent protein, Bacteria; 147336-22-9/Green Fluorescent Proteins; EC 3.4.22.-/CASP3 protein, human; EC 3.4.22.-/CASP8 protein, human; EC 3.4.22.-/CASP9 protein, human; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspase 8; EC 3.4.22.-/Caspase 9; EC 3.4.22.-/Caspases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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