ダウンロード数: 238
このアイテムのファイル:
ファイル | 記述 | サイズ | フォーマット | |
---|---|---|---|---|
journal.pone.0050218.pdf | 3.86 MB | Adobe PDF | 見る/開く |
タイトル: | In Vivo Imaging of Hierarchical Spatiotemporal Activation of Caspase-8 during Apoptosis. |
著者: | Kominami, Katsuya Nagai, Takeharu Sawasaki, Tatsuya Tsujimura, Yuki Yashima, Kenta Sunaga, Yasuhiro Tsuchimochi, Masateru Nishimura, Jun Chiba, Kumiko Nakabayashi, Jun Koyamada, Koji https://orcid.org/0000-0002-4283-3954 (unconfirmed) Endo, Yaeta Yokota, Hideo Miyawaki, Atsushi Manabe, Noboru Sakamaki, Kazuhiro |
著者名の別形: | 酒巻, 和弘 |
発行日: | 21-Nov-2012 |
出版者: | Public Library of Science |
誌名: | PloS one |
巻: | 7 |
号: | 11 |
論文番号: | e50218 |
抄録: | [Background]: Activation of caspases is crucial for the execution of apoptosis. Although the caspase cascade associated with activation of the initiator caspase-8 (CASP8) has been investigated in molecular and biochemical detail, the dynamics of CASP8 activation are not fully understood. [Methodology/Principal Findings]: We have established a biosensor based on fluorescence resonance energy transfer (FRET) for visualizing apoptotic signals associated with CASP8 activation at the single-cell level. Our dual FRET (dual-FRET) system, comprising a triple fusion fluorescent protein, enabled us to simultaneously monitor the activation of CASP8 and its downstream effector, caspase-3 (CASP3) in single live cells. With the dual-FRET-based biosensor, we detected distinct activation patterns of CASP8 and CASP3 in response to various apoptotic stimuli in mammalian cells, resulting in the positive feedback amplification of CASP8 activation. We reproduced these observations by in vitro reconstitution of the cascade, with a recombinant protein mixture that included procaspases. Furthermore, using a plasma membrane-bound FRET-based biosensor, we captured the spatiotemporal dynamics of CASP8 activation by the diffusion process, suggesting the focal activation of CASP8 is sufficient to propagate apoptotic signals through death receptors. [Conclusions]: Our new FRET-based system visualized the activation process of both initiator and effector caspases in a single apoptotic cell and also elucidated the necessity of an amplification loop for full activation of CASP8. |
著作権等: | © 2012 Kominami et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
URI: | http://hdl.handle.net/2433/166071 |
DOI(出版社版): | 10.1371/journal.pone.0050218 |
PubMed ID: | 23185580 |
出現コレクション: | 学術雑誌掲載論文等 |
このリポジトリに保管されているアイテムはすべて著作権により保護されています。