論文 - 詳細
| RRC ID | 38577 |
|---|---|
| 著者 | Sun F, Mikuni S, Kinjo M. |
| タイトル | Monitoring the caspase cascade in single apoptotic cells using a three-color fluorescent protein substrate. |
| ジャーナル | Biochem Biophys Res Commun |
| Abstract |
Fluorescence cross-correlation spectroscopy (FCCS) reveals information about the spatiotemporal coincidence of two spectrally well-defined fluorescent molecules in a small observation area at the level of single-molecule sensitivity. To simultaneously evaluate the activities of caspase-3 and caspase-9, we constructed a chimeral protein that consisted of tandemly fused enhanced cyan fluorescent protein (ECFP), monomeric red fluorescent protein (mCherry) and monomeric yellow fluorescent protein (Venus). In HeLa cell lysates, a combination of tumor necrosis factor-α (TNF-α)- and cycloheximide (CHX-)-induced apoptosis was monitored. In this, decreases of cross-correlation amplitudes were observed between ECFP and mCherry and between mCherry and Venus. Moreover, time-dependent monitoring of single cells revealed decreases in the cross-correlation amplitudes between ECFP and mCherry and between mCherry and Venus before morphologic changes were observed by laser scanning fluorescence microscopy (LSM). Thus, our method could predict the fate of the cell in the early apoptotic stage. |
| 巻・号 | 404(2) |
| ページ | 706-10 |
| 公開日 | 2011-1-14 |
| DOI | 10.1016/j.bbrc.2010.12.047 |
| PII | S0006-291X(10)02285-0 |
| PMID | 21156159 |
| MeSH | Apoptosis* Bacterial Proteins / analysis Bacterial Proteins / genetics Caspase 3 / analysis* Caspase 3 / biosynthesis Caspase 9 / analysis* Caspase 9 / biosynthesis Fluorescent Dyes / analysis Green Fluorescent Proteins / analysis Green Fluorescent Proteins / genetics HeLa Cells Humans Luminescent Proteins / analysis Luminescent Proteins / genetics Recombinant Fusion Proteins / analysis Recombinant Fusion Proteins / genetics Spectrometry, Fluorescence / methods* Tumor Necrosis Factor-alpha / pharmacology |
| IF | 2.985 |
| 引用数 | 4 |
| WOS 分野 | BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY |
| リソース情報 | |
| ヒト・動物細胞 | |