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Simultaneous real-time detection of initiator- and effector-caspase activation by double fluorescence resonance energy transfer analysis.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Kawai, Hiroshi F. Suzuki, Takuo Kobayashi, Tetsu Sakurai, Haruna Ohata, Hisayuki Honda, Kazuo Momose, Kazutaka Namekata, Iyuki Tanaka, Hikaru Shigenobu, Koki Nakamura, Ryu Hayakawa, Takao Kawanishi, Toru |
| Copyright Year | 2005 |
| Abstract | Fluorescence resonance energy transfer (FRET) with green fluorescent protein (GFP) variants has become widely used for biochemical research. In order to expand the choice of fluorescent range in FRET analysis, we designed various color versions of the FRET-based probes for caspase activity, in which the substrate sequence of the caspase was sandwiched by donor and acceptor fluorescent proteins, and studied the potential of these color versions as fluorescent indicators. Six color versions were constructed by a combination of cyan fluorescent protein (CFP), GFP, yellow fluorescent protein (YFP), and DsRed. Real-time monitoring in single cells revealed that all probes could detect caspase activation during tumor necrosis factor (TNF)-alpha-induced cell death as a fluorescent change. GFP-DsRed and YFP-DsRed were as sensitive as CFP-YFP, and CFP-DsRed also showed a large fluorescent change. By using two probes, CFP-DsRed and YFP-DsRed, we carried out simultaneous multi-FRET analysis and revealed that the initiator- and effector-caspases were activated almost simultaneously in TNF-alpha-induced cell death. These findings may give experimental bases for the development of novel techniques to analyze multi-events simultaneously in single cells by using FRET probes in combination. |
| Starting Page | 1219 |
| Ending Page | 1219 |
| Page Count | 1 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | https://www.jstage.jst.go.jp/article/jphs/97/3/97_3_361/_pdf |
| PubMed reference number | 15750288v1 |
| Volume Number | 97 |
| Issue Number | 3 |
| Journal | Journal of pharmacological sciences |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Base CFP gene CFP protocol Caspase Activation Cell Death Cessation of life Cyan Fluorescent Protein Fluorescence Resonance Energy Transfer Green Fluorescent Proteins Numerous Tumor Necrosis Factors Version fluorescent protein 583 |
| Content Type | Text |
| Resource Type | Article |
